Neuroscience Antibodies & Protocols · 2005-11-24 · RID Radial immunodiffusion RIe Rocket immunoelectrophoresis RImm Radial immunodiffusion RipA Radioimmunoprecipitation assay SB

Neuroscience Antibodies & Protocols(U.S.A. & European edition) 2006

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Ordering information: www.abcam.com | Tech support: www.abcam.com/technicalNeuroscience resources: www.abcam.com/neuroscience

ContentsWelcome to Abcam

About Abcam . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1Neuroscience at Abcam . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2

Neuroscience Abwire . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2

Abbreviations

Species key . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5

Neuroscience antibody categories

Cell adhesion proteinsCytoskeletal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6ECM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8Membrane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10Nuclear . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10

Growth & developmentAxonal Guidance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11Neurogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11Neurotrophins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12Notch Pathway . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13Signal Transduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14

NeurodegenerationAlzheimer’s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18Huntington’s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22Parkinson’s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22Prions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .24Related Targets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25

NeuroendocrinologyGH Regulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26Gonadotrophic Axis . . . . . . . . . . . . . . . . . . . . . . . . . . . . .26HPA Axis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27Obesity & Metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . .28

Technical help

Neuroscience resources at Abcam . . . . . . . . . . . . . . . . .64Product datasheet guide . . . . . . . . . . . . . . . . . . . . . . . . .65

Ordering and contact details

Contact details . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .89Ways to order . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .89

Coming soon

New Neuroscience antibodies . . . . . . . . . . . . . . . . . . . . . . .91

Target index

Target index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .92

The team . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2Benefits of an Abcam account . . . . . . . . . . . . . . . . . . . . . . . .3AbreviewsSM and Abcam AbpointsSM . . . . . . . . . . . . . . . . . . . .3

Applications key . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5

Oxytocin & Vasopressin . . . . . . . . . . . . . . . . . . . . . . . . . .30Thyroid Axis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .30

Neuronal markersGlia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .31Neural Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33Neuron . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .35

NeurotransmissionCalcium Signaling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43Intracellular Signaling . . . . . . . . . . . . . . . . . . . . . . . . . . . .44Neurotransmitters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .47Neuropeptides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .50Nitric Oxide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .52Pharmacological Tools . . . . . . . . . . . . . . . . . . . . . . . . . . .53Receptors & Channels . . . . . . . . . . . . . . . . . . . . . . . . . . .53Secretory Vesicles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .59Transporters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .61

Sensory pathwaysHearing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .62Nociception . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .62Olfaction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .63Touch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .63Vision . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .63

Neuroscience protocols and tips . . . . . . . . . . . . . . . . . . .66Neuroscience signaling pathway diagrams . . . . . . . . . . .85

Required Information for ordering . . . . . . . . . . . . . . . . . . . .89Abcam’s terms and conditions . . . . . . . . . . . . . . . . . . . . . . .90

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Customer ServiceTel: Europe +44 (0) 1223 696000 | U.S.A. 1 617 225 2272 (Toll free: 1 888 77 ABCAM)

About Abcam

Abcam, located in Cambridge, U.K. and Cambridge, MA, U.S.A., was founded in 1998 by Dr. JonathanMilner, then a researcher at Cambridge University, U.K.:

“Like many life science researchers, I was frustrated by the time it took to locate and selectantibodies essential for my research. This was largely due to poor information and out-of-datecatalogs from the vast range of suppliers who were spread across many countries. In some cases, Ialso experienced difficulties with companies whose products were unreliable and whose customerservice was slow and unhelpful. My vision was to build a company that offered reliable cutting-edgeproducts and great customer service.”

It was a tough vision - Abcam was created to sell the best antibodies in the world with the most comprehensive, honest andup-to-date datasheets, fast delivery, helpful customer service and comprehensive technical support.

Seven years on and with thousands of products across our range, Abcam ships toscientists in over 55 countries around the world. Abcam is committed to thefollowing fundamental principles that have seen us become the first choicefor thousands of life-scientists throughout industry and academia:

• Quality of products and datasheets• Helpful technical support and online technical resources• Friendly customer service • Fast delivery

Comprehensive

We have a policy of honesty - our customers know everything that we know aboutour products, as soon as we know it. Everything we know about each product isposted on the datasheet. To achieve exceptional levels of quality for our customerswe provide:

• Comprehensive datasheets. • Customers’ AbreviewsSM of our products .• Product-specific protocols used by our customers.• Customers’ technical questions and our answers for each product.• Product information and new products updated daily.

Helpful

Our Technical Support and Customer Service departments are full of scientists that have many years of experience workingwith the types of products that we supply to our customers.We provide continuous order progress updates, so that you always know what is happening with your order and when youshould expect to receive it.

The Queen's Award for Enterprise

The Queen's Award for Enterprise has been running since 1966 and is widely recognised as the premieraward for business performance in the UK; granted annually for outstanding business achievement.Abcam demonstrated excellent year-on-year growth in overseas sales between 2002 and 2004 with 84%of sales during these three years coming from overseas customers. The award is positive proof of thecompany's outstanding success as a customer-focused international business.

Welcome to Abcam

...all at www.abcam.com!

Free services

The World’s Antibody Gateway search engineIf we do not have the product that you are looking for use this service to search all online catalogs of other antibody suppliers.Find the “WAG” at:www.abcam.com/antibodygateway

The Abcam ToolbarDirectly search your choice of Abcam, GenBank, Google, LocusLink, OMIM, PubMed, SWISSProt. The toolbar sits within yourinternet browser, so that you can use it from wherever you are on the internet. Get this useful facility at:www.abcam.com/toolbar

Fast

We have developed web-based systems that enable us to quickly identify cutting-edge products, make them rapidly availableto our customers and then deliver them by Express Delivery.

We are pleased to be able to offer next-day delivery to all major cities as well as same day delivery to customers in CambridgeU.K. and Cambridge MA, U.S.A.

Neuroscience at Abcam

Our range of over 2,000 Neuroscience primary antibodies has a breadth and depth that is hard to beat. Further more, wecollaborate with leading academics to make the Abcam range one of the best in the world!

The Neuroscience Abwire Our Neuroscience Abwire web page is full of exclusive articles, product browsing tools, conference and publication informationand specialist protocols. Visit today for cup-to-date information in this rapidly progressing field.

Look out for new Neuronal Marker IHC Galleries, Neurodegeneration and Neurotransmission signaling pathway diagrams andNeuroscience Product Locator tools (to name but a few).

Visit: www.abcam.com/neuroscience for more.

The Neuroscience teamAbcam has a specialized team working on sourcing and testing the best Neuroscience antibodies available. Our background inthe neurosciences is very strong to better serve our customers. Members include:

Technical SupportSarah Mardle (PhD King’s College, University of London, U.K.) Technical Support Manager (U.K.)

Bill Campbell (PhD Brown University, U.S.A.)Technical Support Manager (U.S.A.)

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Sarah and Bill help customerswho have enquiries about, orproblems with, Abcamantibodies using their extensivePhD and postdoctoral laboratoryexperience.They also:

• Attend conferences to keep up-to-date with antibody techniques and research

• Source new protein targets

• Source novel techniques using antibodies

• Build collaborations with academics.

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Neuroscience at Abcam

The Neuroscience team

Business DevelopmentSabrina Edmonds (PhD University of Cambridge, U.K.)Business Development Manager for Neuroscience

Sabrina is responsible for sourcing new and exciting protein targets to generate antibodies against byattending conferences, forging collaborations with academics and by reading current literature. She isthe main contact for academics who are interested in licensing their neuroscience antibodies for salethrough Abcam.

MarketingRhian Hayward (PhD University of Oxford, U.K.)Neuroscience Marketing Manager

Rhian is responsible for marketing Abcam’s Neuroscience range to our customers worldwide. Sheensures that Abcam supports many Neuroscience community activities through conferencesponsorships, informative marketing materials e.g. signalling pathways (see page 83), theNeuroscience Abwire and more.

Suggestions or comments: Email the team at [email protected]

Benefits of an Abcam Account

Take full advantage of all our online Neuroscience resources by opening an account.• Earn Abcam AbpointsSM

• Buy quickly and efficiently online.• Customise your antibody updates. We can notify you by email of new and updated antibodies related directly to the

protein families which interest you.

AbreviewsSM

Our unique system for enabling our customers to feedback to you and us about the performance of our primary antibodies. It isimportant to know how our antibodies perform under different conditions and that they are working as expected. We encouragethis feedback (positive and negative) by awarding AbpointsSM for each Abreview:

Customers’ Abreviews are posted on the product datasheets for others to browse. You can contact the reviewer to ask furtherquestions too!

AbreviewTM Action Points Description

Submit an Abreview 50 Each unique combination of antibody, application and sample species that youreview, earns you Abcam AbpointsSM, e.g. if you use one of our antibodies inWB on Human and Mouse samples and submit an Abreview for both thesecombinations, then you will receive 50 Points for each one.

Submit an Abreview(untested application /species)

200 As for “Submit an Abreview” but for an antibody that has not yet been tested inthat particular species or application.

Submit an image with yourAbreview

100 If you submit an image to accompany any of your Abreviews, then you willreceive extra Points when it is published with your Abreview. For Abreview andimage publication criteria, please see the Abreviews section of our website.

Fast Track Abreviewsbonus

950 Be the 1st researcher to submit conclusive positive or negative data (throughAbreviews see below) about one of our Fast Track antibodies and earn this bonus(in addition to the Abcam Points that you will earn for submitting your Abreview).

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Abcam AbpointsSM

In addition to AbreviewsSM, you can also earn AbpointsSM by buying your antibodies online at www.abcam.com or by registeringyour interests there.

AbpointsSM rewards• Vouchers for leading retailers:

• Useful gifts:

• Discounts on Abcam products:

• Money to go to conferences / for journal subscriptions:

• Donate money to charities: ...You can even suggest new rewards.

Collecting and spendingRead about collecting and spending AbpointsSM and get the complete list of rewards for your country at: www.abcam.com/abpoints.

Check out thelatest at

www.abcam.com

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Key Species

Many wide range of species

At Arabidopsis thalianaAm AmphibianAv AvianBa BaboonBu Burkholderia spp.Ca CatCe Caenorhabditis elegansCh ChickenC CowDe DeerD DogDm Drosophila melanogasterEl ElkEc Escherichia coli

Key Species

Fe FerretFi FishGt GoatG Goose Gp Guinea pigHa Hamster H HorseHu HumanIn InsectK KangarooLi LizardMm MouseMa MammalsMi MinkMk MonkeyOp Opossum

Key Species

Pi PigP PlantsQ QuailRb RabbitRt RatRe ReptilesRo RodentsSc Sacchromyces cerevisiaeS SalamanderSha SharkSh SheepSn SnakeX Xenopus laevisY all Yeast

Abbreviations


Key Applications

AF Acetone fixedAgg AgglutinationAI Adhesion inhibition AP Affinity purificationBL BlockingCCIe Counter current

immunoelectrophoresisChIP Chromatin immunoprecipitationConjugation ConjugationDID Double immunodiffusionDot Dot blotEC Experimental controlEIA Enzyme immunoassayELISA ELISAEM Electron microscopyEMSA Electrophoretic mobility shift assayFACS Fluorescence activated cell sortingFC Flow cytometryFuncS Functional studiesGSA Gel supershift assaysH HistologyI-ELISA Indirect ELISAIA ImmunoassayIB ImmunoblottingICC ImmunocytochemistryID ImmunodiffusionIe ImmunoelectrophoresisIF ImmunofluorescenceIHC ImmunohistochemistryIHC-F Immunohistochemistry (Formalin fixed sections)IHC-Fr Immunohistochemistry (Frozen sections)

Key Applications

IHC-P Immunohistochemistry (Formalin-fixed paraffin-embedded sections)

IM ImmunomicroscopyInhib Inhibition assayIP ImmunoprecipitationIRMA Immunoradiometric assayISH In situ hybridizationMct Microcytotoxicity testingNeut NeutralizingP PurificationPA Prothrombin assayPCC Primary cell culturePCR Polymerase chain reactionRIA RadioimmunoassayRID Radial immunodiffusionRIe Rocket immunoelectrophoresisRImm Radial immunodiffusionRipA Radioimmunoprecipitation assaySB Southern blotTCC Transfected cell cultureTLC Thin layer chromatographyWB Western blotting

OtherFast Track Application and species data pendingPep. avail. Peptide available

AbbreviationsSpecies key

Applications key

Many of the images in this catalog have been supplied by Abcam’s collaborators in the Neuroscience field. We wish to thank allthese contributors and we look forward to working with you in the future!

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Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Intermediate Filaments

Target Clonality Applications tested Host Species reactivity Datasheetwww.abcam.com/...

GARP (truncated) P WB Rb Rt, B ab15094GFAP P IF, IHC-Fr Ch Hu, Mm, Rt, F, C ab4674GFAP P IF, IHC-F, IHC-P, IP, PCC, WB Rb F, B, Hu, Mm, Pi, Rt ab7260GFAP P IF, IHC-F, IHC-Fr, IHC-P Rb Hu, Rt, B ab7779GFAP [2A5] M IF, IHC-F, IHC-P, PCC, WB Mm Ma ab8136GFAP [6F2] M IHC-Fr, IHC-P, IM,

TEM immunogold labeling, WB Mm Hu ab8975GFAP [GF-01] M ICC, IHC-Fr, IHC-P, IP, WB Mm Hu, F, Pi

Does not react with Mm or Rt ab7806GFAP (phospho T7) [YC10] M ICC, WB Mm Hu, C, Pi

Does not react with Mm or Rt ab12340Internexin alpha P ICC, IHC-Fr, WB Rb Hu, Mm, Rt, C, F ab22038Internexin alpha [1D2] M ICC, IHC-P, WB Mm Hu, Rt ab22039Internexin alpha [2E4] M ICC, IHC-Fr, IHC-P, WB Mm Hu, Rt ab22040Laminin P ELISA, IF, IHC-P, RIA Rb Hu ab23753Laminin P ELISA, WB Ch Hu, Mm, Rt ab14055Laminin P Dot, IF, IHC-F, IHC-P Rb Ma, Am, Av, Re ab11575Laminin [A5] M ICC, IHC-Fr, WB Rt Hu, Mm, Ba ab2500Laminin [LAM-89] M IF, IHC-P Mm Hu, F, Rb.

Does not react with Mm, Rt, Ch, D,Gp, Li, Pi, Sh or Sn ab11574

Laminin [MEC5] M IHC-Fr Rt Hu, Mm, Rt ab2466Laminin 2 alpha [4H8-2] M ELISA, ICC, IF, IP, WB Rt Hu, Mm ab11576Laminin 5 P IHC-Fr, WB Rb Hu ab14509Laminin B2 gamma 1 [SPM277] M IF, IHC-P, IP, WB Rt Hu, Mm ab17792Laminin S [C4] M IP, WB Mm Hu, Rt, B, Pi, Gp, Ch ab328967kDa Laminin Receptor P ICC, IF, IHC-P, IP, WB Rb Hu, Mm ab71167kDa Laminin Receptor [MLuC5] M FACS, ICC, IF, IHC-P Mm Hu, Rt ab3099Nestin P ICC, IF Rb Hu, Ha, Mk

Does not react with Mm ab5968Nestin [10C2] M IF, IHC-Fr, IHC-P, WB Mm Hu.

Does not react with rodent nestin ab22035Nestin [2C13A11] M FACS, ICC, IP, WB Mm Hu.

Does not react with Rt ab18102Nestin [2Q178] M EM, ICC, IHC-Fr, IHC-P, WB Mm Mm, Rt

Does not react with Hu, F, Mk or Sh ab6142Nestin [3k1] M ELISA, FACS, ICC,

IHC (aldehyde-based fixation), IP, WB Mm Hu ab632068kDa Neurofilament [1D2] M ICC, IF, IHC-Fr, IHC-P, WB Mm Hu, Rt ab457168kDa Neurofilament [NR4] M IF Mm Hu, Rt, Ch, Pi ab1121968kDa Neurofilament [SPM204] M IHC-P, WB Mm Hu ab1783270 kDa Neurofilament Light P IHC-F, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, B, Pi, Av ab903570 kDa Neurofilament Light [2F11] M IHC-Fr, IHC-P, WB Mm Hu, Op ab897070 kDa Neurofilament Light [DA2] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt, Ch, C, Pi ab4572160 kDa Neurofilament Medium P IF, IHC-F, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, F, B, Pi, Re, Av ab9034160 kDa Neurofilament Medium [3H11] M IHC-F, IHC-Fr, IHC-P, WB Mm Hu ab7256160 kDa Neurofilament Medium [RNF403] M IHC-Fr, WB Mm Hu, Rt ab9271

160 kD Neurofilament Medium antibody [NF-09] (ab7794)

IF - ab7794 at a dilution of 1/1000, staining on rat brain tissue (30µm thick coronal sections) in free floating IHC.

Clonality Applications tested Host Species reactivity DatasheetM WB, ELISA, IHC-P, IHC-Fr, IF, ICC Mm Ma www.abcam.com/ab7794

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200 kDa Neurofilament Heavy P IF, IHC-Fr, IHC-P, WB Rb F, B, Hu, Mm, Pi, Rt ab8135200 kDa Neurofilament Heavy [NAP4] M ELISA, IHC-F, IHC-Fr, IHC-P, WB Mm Av, Ma ab7257200 kDa Neurofilament Heavy [RNF402] M IHC-Fr, IHC-P, WB Mm Hu, Rb, Rt, Mm, B, D, Sh,

Gp, Ch, Ha, Mk, X ab3966200 kDa Neurofilament Heavy [SPM203] M IHC-P, WB Mm Hu ab17879200kDa + 70kDa Neurofilament [2F11] M IF, IHC, IHC-P, IP, WB Mm Hu ab17114Peripherin P ELISA, ICC, IF, IHC-F, IHC-P, WB Rb Ma ab7258Peripherin [2Q135] M ICC, WB Mm Hu, Mm, Rt, C, Pi.

Does not react with Ch ab17999Peripherin [7C5] M ICC, IHC-P, WB Mm Ch ab4573Peripherin [8G2] M ELISA, ICC, IHC-F, IHC-P, WB Mm Ma ab7657Peripherin variant PER56 P IHC-Fr, IHC-P, WB Rb Hu, Mm ab4655Peripherin variant PER61 P IHC-Fr, IHC-P, WB Rb Hu, Mm ab4646

Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Microtubules


beta III Tubulin [TU-20] M ICC, IF, IHC-P, IP, WB Mm Ma ab7751beta III Tubulin [TUJ-1] M ICC, IF, IHC-Fr, WB Mm Hu, Rt ab14545Doublecortin P IHC-Fr Gp Hu, Mm, Rt ab10293MAP2 P IF, IHC, WB Ch B, Hu, Mm, Rt ab5392MAP2 [AP-20] M ICC, WB Mm Mm, Rt, X, B, Q, S ab11268MAP2 [HM-2] M ICC, IHC, WB Mm Hu, Mm, Rt, B, Ch, Q ab11267MAP2 [MT-01] M ICC, IF, IP, WB Mm Mm, F, C, Pi ab7756MAP2a + MAP2b [AP20] M ICC, IF, IHC-P, WB Mm Hu, Rt, Mm, B, Ch, X , Q ab3096MAP2a + MAP2b [MT-01] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, F ab8479MAP5 [3G5] M IF, IHC-P, IHC on primary cultures Mm Hu, Mm, Rt, C.

Does not react with Ch ab3095

Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Regulation


Thrombospondin 1 [CSI 002-65] M ELISA, IHC, IHC-Fr, IP Mm Hu, B ab23420

Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Thin Filaments


MYRIP P Fast track Gt Fast track ab10149 Pep. avail.

200 kD Neurofilament Heavy antibody [NF-01] (ab7795)

IF - ab7795 at a dilution of 1/1000, staining in rat brain tissue (30 µm thick coronal sections) on free floating IHC.

Clonality Applications tested Host Species reactivity DatasheetM WB, ELISA, IHC-P, IHC-Fr, IF, ICC Mm Ma www.abcam.com/ab7795

Cell adhesion proteins


Neuroscience > Cell Adhesion Proteins > Cytoskeletal Proteins > Intermediate Filaments


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Neuroscience > Cell Adhesion Proteins > ECM Proteins


Agrin [AGR 131] M ICC, IHC, IRMA, WB Mm Mm, Rt ab12362Agrin [Agr 247] M ICC, IF Mm Mm, Rt ab12364Agrin [Agr 86] M FACS, ICC, WB Mm Mm, Rt ab12361Amisyn P Fast track Gt Fast track ab5899 Pep. avail.Bestrophin P ICC, IHC, IP, WB Rb Hu, Mm, Rt ab14927Bestrophin M WB Mm Hu, Pi, Mk ab2182Drebrin P WB Rb Rt, Hu ab11068Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350Fetuin A P ELISA, WB Rb Hu ab13980Laminin P ELISA, IF, IHC-P, RIA Rb Hu ab23753Laminin P ELISA, WB Ch Hu, Mm, Rt ab14055Laminin P Dot, IF, IHC-F, IHC-P Rb Ma, Am, Av, Re ab11575Laminin [A5] M ICC, IHC-Fr, WB Rt Hu, Mm, Ba ab2500Laminin [LAM-89] M IF, IHC-P Mm Hu, F, Rb.

Does not react with: Mm, Rt, Ch, D, Gp, Li, Pi, Sh or Sn ab11574

Laminin [MEC5] M IHC-Fr Rt Hu, Mm, Rt ab2466Laminin 2 alpha [4H8-2] M ELISA, ICC, IF, IP, WB Rt Hu, Mm ab11576Laminin 5 P IHC-Fr, WB Rb Hu ab14509Laminin B2 gamma 1 [SPM277] M IF, IHC-P, IP, WB Rt Hu, Mm ab17792Laminin S [C4] M IP, WB Mm Hu, Rt, B, Pi, Gp, Ch ab328967kDa Laminin Receptor P ICC, IF, IHC-P, IP, WB Rb Hu, Mm ab71167kDa Laminin Receptor [MLuC5] M FACS, ICC, IF, IHC-P Mm Hu, Rt ab3099Mint3 P WB Rb Rt, Mm ab3450 Pep. avail.NCAM [123C3] M IF, IHC-F, IHC-Fr, IHC-P Mm Rt, Hu ab8077NCAM [B332 (123A8)] M FACS, IHC-Fr, WB Mm Hu ab8596NCAM [ERIC-1] M ELISA, IHC-Fr, IP, RIA, WB Mm Hu ab6123NCAM [H28-123] M AP, IHC-Fr, IM, WB Rt Mm ab19782NCAM [MEM-188] M FACS, IP, WB Mm Hu ab8233NCAM [RNL-1] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt ab9018Nicastrin P WB Rb Hu, Mm ab3444 Pep. avail.SIRP [MRC OX-41 ] M FACS, IHC-Fr, IHC-P, WB Mm Rt ab9295SIRP alpha P IHC-P, WB Rb Hu, Mk.

Does not react with Mm ab2971SIRP alpha P WB Rb Hu, Rt, Mm ab8120 Pep. avail.

Reelin antibody [142] (ab18569)

IHC-P - ab18569 detecting Reelin immunoreactivity in Cajal-Retzius cells of the mouse cortex at postnatal day 0.ab18569 was used at 1/1000 (overnight incubation at 4°C). Immunoreactivity was visualized using ABC amplification.

Clonality Applications tested Host Species reactivity DatasheetM ELISA, ICC, IHC-P, IP Mm Many www.abcam.com/ab18569

67kD Laminin Receptor antibody (ab2508)

IF - staining observed in the hippocampus in the cytoplasm of some astrocytes and neurons using ab2508.Recommended dilution of antibody is 1:300-1:1000 with overnight incubation at room temperature. Free floating IHCperformed with TSA amplification.

Clonality Applications tested Host Species reactivity DatasheetP IHC-Fr, Fl, IP, WB Rb Hu www.abcam.com/ab2508

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Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260Synaptotagmin P IF, WB Rb Hu, Rt ab10104Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259Synaptotagmin [ASV48] M ICC, IHC, IP, WB Mm Rt, B, Mk, Sha, Mm ab12255SynCAM P WB Rb Rt ab3910Syntrophin [1351] M IF, IP, WB Mm Hu, Mm, Rt, Am, Ch, D, Fi ab11425Syntrophin gamma 2 P ELISA, WB Gt Hu ab15713 Pep. avail.Tenascin C P WB Ch Hu ab16290Tenascin C [MTn-12] M AF, Dot, ELISA, IF, IHC, IHC-F,

IHC-Fr, IP, WB Rt Mm ab6346

Reelin antibody [G10] (ab18570)

IHC-P - ab18570 detecting Reelin immunoreactivity in Cajal-Retzius cells of the mouse cortex at postnatal day 0.ab18569 was used at 1/5000 (overnight incubation at 4°C). Immunoreactivity was visualized using ABC amplification.NB: Antigen retrieval performed (microwave in citrate buffer pH 6).

Clonality Applications tested Host Species reactivity DatasheetM WB, IP, ELISA, IHC-Fr, ICC, IHC-P Mm Mm, Rt. Does not react with Hu www.abcam.com/ab18570 C

ell adhesion proteins


Neuroscience > Cell Adhesion Proteins > ECM Proteins


NeuroscienceAbwire at

www.abcam.com/neuroscience

10

Neuroscience > Cell Adhesion Proteins > Membrane Proteins


Cathepsin B P ID, WB Sh Hu ab211Cathepsin B P IHC-Fr, IHC-P Rb Hu ab7670Cathepsin B [3E1] M IHC-Fr, IHC-P Mm Hu ab7430Cathepsin D P ELISA, WB Rb Hu ab19555Cathepsin D P IHC-Fr, IHC-P Rb Hu ab826Cathepsin D [CTD-19] M ELISA, IHC-F, IHC-P, WB Mm Hu ab6313Cathepsin D [D101] M IHC-Fr, IHC-P Mm Hu ab7433Cathepsin G P ELISA, WB Rb Hu ab20725Cathepsin G P IHC-Fr, IHC-P Rb Hu ab15988Cathepsin G P WB Sh Hu ab8816Cathepsin H [1D10] M IHC-Fr, IHC-P, WB Mm Hu ab7432Cathepsin L P IHC-Fr, IHC-P Sh Hu ab7454Cathepsin L [33/2] M ELISA, IHC-Fr, WB Mm Hu, Mm, Rt, Mi ab6314Heparan Sulfate Proteoglycan 2 [CSI 001-74] M AP, ELISA, IHC, IHC-Fr, IP, WB Mm Hu, C ab23418Heparan Sulfate Proteoglycan 2[SPM255], prediluted M IHC-P Rt Hu, Mm, B, Pi ab17848MOSP [CE1] M IF, IHC-Fr Mm Hu, Rt, Mm, F, Ch, Mk, Ca ab3094Myelin Basic Protein P WB Rb Hu, Mm, Rt ab12355Myelin Basic Protein P IF, IHC-Fr, IHC-P Rb Hu, Rt ab2404Myelin Basic Protein [2B5] M ICC, IHC-F, IHC-Fr, IHC-P Mm Hu ab8053Myelin Basic Protein [12] M AF, ELISA, IF, IHC-Fr, PCC,

RIA, WB Rt Hu, B, Gp, Mm, Rb, Rt, Sh ab7349Myelin Basic Protein [26] M IHC-Fr Mm MBP from most species ab11162Myelin Basic Protein [B505] M IHC-Fr, IHC-P Mm Hu, Rb, Rt, C, Mk ab8764Myelin Basic Protein [Clone 2] M ELISA, IHC-Fr Mm Hu, Rt, B, Gp, Rb, Sh ab11159Myelin PLP [plpc 1] M IF, IHC-Fr, WB Mm Ma ab9311PMP22 P IHC-P Rb Hu ab15506PMP22, prediluted P IHC-P Rb Hu ab15507PMP22 [20-14] M IHC-P Mm Hu ab3278SORL1 P Fast track Gt Fast track ab22814Spinesin P WB Gt Mm ab2245

ProteinsMyelin Basic Protein protein (Biotin) ab792

Neuroscience > Cell Adhesion Proteins > Nuclear Proteins


Galectin 3 P IHC, WB Rb Hu ab2473Galectin 3 [A3A12] M ELISA, FACS, IF, IHC-Fr, IP, WB Mm Hu, Rt, Mm ab2785Galectin 7 P IP, WB Rb Mm ab10482

NrCAM antibody (ab24344)

IF - ab24344 (1/300) detects NrCAM on embryonic day 11.5 mouse spinal cord. Floor plate commissurral axonscrossing to the contralateral side of the spinal cord are shown here labeled for NrCAM. Mouse spinal cord was 4%PFA-fixed and processed on slide for IF.

Clonality Applications tested Host Species reactivity DatasheetP IF, IHC-F, IHC-P, IHC-FrFl, IP, WB Rb Hu, Mm, Rt www.abcam.com/ab24344

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Neuroscience > Growth and Development > Axonal Guidance Proteins


Aggrecan ARGxx [BC-3] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, B, F, D, Gp, H, Pi,Rb, Rt, Sh ab3773

AMIGO2 P WB Rb Mm, Rt ab16762CRMP5 [CR-1] M IHC-Fr, WB Rt Hu, Rt, B ab19378CRMP5 [CR-3] M IHC-Fr, WB Rt Hu, Mm, Rt, B ab19352EphA1 P ELISA, WB Rb Hu ab5385EphA1 P I-ELISA, WB Gt Hu ab7036EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387EphA2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610EphA3 P I-ELISA, WB Gt Mm ab7038EphA3 P ELISA, WB Gt Hu.

Does not react with Mm ab10611EphA3 [III-A4] M FACS, IP Mm Hu ab19439EphA4 P ELISA, WB Rb Hu ab5389EphA4 P I-ELISA, WB Gt Mm ab7039EphA4 P I-ELISA, WB Gt Hu ab7041EphA5 P ELISA, WB Rb Hu, Mm ab5397EphA5 P ELISA, WB Gt Mm, Rt ab10612EphA6 P ELISA, WB Gt Mm ab10613EphA7 P ELISA, WB Rb Hu ab5411EphA7 P ELISA, WB Gt Mm ab10614EphA8 P ELISA, WB Gt Mm ab10615EphB1 P ELISA, WB Rb Hu, Mm ab5414EphB1 P I-ELISA, WB Gt Rt ab7042EphB1 P Dot, IP Sh Hu ab10406EphB2 P ELISA, WB Rb Hu, Mm ab5418EphB2 P Dot, ELISA Gt Mm ab10616EphB3 P ELISA, WB Gt Mm ab10617EphB3 P ELISA, WB Gt Hu ab7044EphB4 P Dot, ELISA, Inhib Gt Mm ab10618EphB6 P Dot, ELISA Gt Mm ab10619L1CAM [UJ127] M IF, IHC-P, IP, WB Mm Hu ab3200L1CAM [UJ12711] M IP, WB Mm Hu ab20148L1CAM [UJ1814] M IP, WB Mm Hu ab20149Neuropilin 1 P ELISA, WB Gt Hu ab18130Neuropilin 1 P IHC-P, WB Rb Rt, Mm ab16786Neuroserpin P IHC, WB Rb Hu ab16171NRG1 P WB Rb Hu, Mm ab2994NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369NRP2 P WB Ch Hu ab15874Reelin [142] M ELISA, ICC, IHC-P, IP, WB Mm Ma, Av ab18569Reelin [G10] M ELISA, ICC, IHC-Fr, IHC-P, IP, WB Mm Mm, Rt.

Does not react with Hu ab18570Robo1 P ELISA, IF, WB Rb Mm ab7279Robo4 P WB Rb Mm ab10547Slit1 P WB Rb Mm, Rt ab10984Slit2 P Fast track Rb Fast track ab7665

ProteinsRobo4 protein ab19112

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Neuroscience > Growth and Development > Neurogenesis


Axotrophin P WB Ch Hu, Mm, Rt ab13997Cdk5 P ELISA, WB Rb Hu, Mm, Rt ab19426Cdk5 [SPM236], prediluted M IHC-P Mm Hu, Mm, Rt ab17875DYRK1A P IF, IP, WB Sh Hu ab16140PTPR P WB Rb Hu ab12153STMN2 P Fast track Gt Fast track ab21190

Neuroscience > Growth & Development > Neurotrophins


Artemin P WB Rb Hu ab1136BDNF P ELISA, WB Rb Hu ab9793BDNF [35928.11] M ELISA, WB Mm Hu ab10505BDNF (Biotin) P ELISA, WB Rb Hu ab12385BMP2 P Dot, ELISA, WB Rb Hu, Mm, Rt ab14933BMP2 P ELISA, WB Rb Hu ab17885BMP2 [65529.111] M I-ELISA, WB Mm Hu ab6285BMP2 (Biotin) P ELISA, WB Rb Hu ab17597CNTF P I-ELISA, Neut, WB Gt Rt ab10834CNTF P ELISA, Neut, WB Rb Rt ab9785CNTF P ELISA, IHC, WB Ch Hu, Mm, Rt ab17692CNTF [AS23] M ELISA, IP, WB Mm Hu ab17284CNTF (Biotin) P ELISA, WB Rb Rt ab12426GDNF P I-ELISA, Neut, WB Gt Hu ab10835GDNF P Dot, IHC-Fr, WB Sh Hu ab6206GDNF P ELISA, IHC, WB Ch Hu, Mm, Rt ab17637GDNF family Receptor alpha 2 P IHC-Fr, WB Rb Mm, Rt ab6170GDNF family Receptor alpha 3 P IHC-Fr, WB Rb Mm, Rt ab6171GDNF family Receptor alpha 3 P WB Rb Hu, Rt, Mm ab8028 Pep. avail.GDNFR alpha P WB Rb Hu, Rt, Mm ab8026 Pep. avail.GFR alpha 2 P WB Rb Hu, Mm, Rt ab19146

BDNF antibody (ab6201)

IHC - immunostaining BDNF-containing hippocampal neurons in coronal rat brain sections (30 microns). ab6201 usedat 1/100 dilution.

Clonality Applications tested Host Species reactivity DatasheetP Dot, ELISA, IHC-Fr, Neut, WB Rb Hu, Rt www.abcam.com/ab6201

FOX1A antibody (ab12161)

ICC - ab12161 staining (1/200) in mouse E11 cortical cells cultured for 6 days.

Clonality Applications tested Host Species reactivity DatasheetP WB, IF, ICC, IHC Rb Hu, Mm www.abcam.com/ab12161

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GFR alpha 3 P WB Rb Hu, Mm, Rt ab19135LANGF Receptor [MC-192] M FACS, IHC-Fr Mm Rt ab6172Neurotrophin 3 P Dot, ELISA, IHC-Fr, Neut, WB Rb Hu ab6202Neurotrophin 3 P ELISA, IHC-Fr, IHC-P, WB Gt Hu, Rt ab10334Neurotrophin 3 (Biotin) P ELISA, WB Gt Hu ab17524Neurotrophin 4 P ELISA, Neut, WB Gt Hu ab9824Neurotrophin 4 M ELISA Mm Hu ab9330Neurotrophin 4 (Biotin) P ELISA, WB Rb Hu ab17888Neurotrophin 4/5 P Dot, ELISA, IHC-Fr, Neut, WB Rb Hu ab6205Neurturin P Dot, ELISA, IHC-Fr, WB Sh Hu ab6208Neurturin P WB Rb Hu ab8061Neurturin peptide BL ab8393NGF P Inhib, Dot, ELISA, IHC-Fr, Neut, WB Rb Mm ab6198NGF [1.A.15] M ELISA Mm Hu ab18167NGF 2.5S P Dot, Inhib, WB Rb Hu, Mm, Ch ab10515NGF beta P ELISA, Neut, WB Gt Hu, Mm, Rt ab10837NGF beta [25623.1] M Dot, ELISA, Inhib Mm ab10513NRG1 P WB Rb Hu, Mm ab2994NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369p75 NGF Receptor P Dot, FACS, IF, IHC-Fr, IHC-P, IP, WB Rb Mm, Rt ab8874p75 NGF Receptor [12A8] M Dot, ICC, IHC-Fr, IP, WB Rt Mm ab8876p75 NGF Receptor [ME20.4] M EM, ELISA, FACS, ICC, IF, Mm Hu, F, Pi .

IHC-P, Inhib, IP, RIA, WB Does not react with Mm, Rt or Ch ab10495p75 NGF Receptor [NGFR5] M FACS, IF, IHC-P Mm Hu, Ba, F, Ca, Mk, Rb

Does not react with Mm or Rt ab3125Pleiotrophin P ELISA, WB Rb Hu ab14025Pleiotrophin P ELISA, WB Gt Hu ab10849SPON1 P WB Ch Hu, Mm, Rt, C ab14271TrkA (phospho Y490) P ICC, IP, WB Rb Hu, Mm, Rt ab1445TrkA/B [1.A.37] M IP, WB Mm Hu ab18137TrkB P IHC-Fr Rb Rt ab6180TrkB P I-ELISA, WB Gt Hu ab10841

ProteinsBDNF protein ab9794CNTF protein ab9786GDNF protein ab9790Neurotrophin 3 protein ab9792Neurotrophin 4 protein ab9825Neurturin protein ab9840NGF beta protein ab9796

TrkA antibody (ab8871)

IF - ab8871 immunostaining in terminals of primary afferent fibers (fibers in the top part of the figure) of the dorsal rootganglion. Free floating IHC performed on 30um cryostat sections. Primary antibody ab8871 was used at 1/3000 andincubated overnight at room temperature.

Clonality Applications tested Host Species reactivity DatasheetP IHC-Fr, IHC-FrFl, IF Rb Rt www.abcam.com/ab8871

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Neuroscience > Growth & Development > Neurotrophins


14

Neuroscience > Growth and Development > Notch Pathway


ADAM10 P WB Rb Hu ab10956APH1a P WB Rb Rt, Hu, Mm ab12104 Pep. avail.APH1a P IF, IP, WB Rb Hu, Mm ab19390DLL4 P ELISA, IHC-F, IHC-P, WB Rb Hu, Mm ab7280Jagged 1 P ELISA, FACS, Neut, WB Gt Hu, Rt ab10580Jagged 2 P Fast track Rb Fast track ab10556LNX P Fast track Gt Fast track ab4538MAML1 P Fast track Gt Fast track ab17019 Pep. avail.Notch1 [A6] M FACS, IHC-P, WB Mm Hu, Mm ab3294Notch1 (activated) P Co-IP, ICC, IF, IP, WB Rb Hu, Mm ab8925 Pep. avail.Notch2 (activated) P Fast track Rb Fast track ab8926 Pep. avail.Notch1 intra P WB Rb Hu.

Does not react with Mm ab8387 Pep. avail.Notch1 [SPM206], prediluted M IHC-P Mm Hu, Mm ab17843Notch1 intra + Notch2 intra P Fast track Rb Fast track ab8928 Pep. avail.Notch 2 intra P WB Rb Hu ab8927 Pep. avail.NUMB P IF, IHC-P, IM, WB Gt Hu, Mm ab4147 Pep. avail.NUMB P IP, WB Rb Hu, Mm ab14140 Pep. avail.NUMBL P Fast track Gt Fast track ab5641 Pep. avail.PEN2 P IP, WB Rb Hu ab16555Presenilin 1 P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab7297Presenilin 1, prediluted P IHC-P Rb Hu ab15508Presenilin 1 [APS 11] M ELISA, IF, IHC-P, WB Mm Hu, Mm, Rt ab15456Presenilin 1 [APS 18] M ELISA, IF, WB Mm Hu, Mm, Rt ab15458Presenilin 2 P ELISA, IHC-Fr, IHC-P Gt Hu ab11148Presenilin 2 P ELISA, IHC, WB Ch Hu, Mm, Rt ab18041Presenilin 2 [198C67921] M WB Mm Hu ab13926Presenilin 2 [APS 26] M ELISA, IF, WB Mm Hu, Mm ab15549SIAH Interacting Protein P Fast track Gt Fast track ab4276 Pep. avail.SIAH1 P WB Gt Hu ab2237 Pep. avail.

Neuroscience > Growth and Development > Signal Transduction


14-3-3 P IP, WB Rb Hu, Mm, Rt ab906314-3-3 P IHC-P, IP, WB Rb Hu, Mm, Rt ab608114-3-3 [2Q248] M WB Mm Rt ab1412114-3-3 [3F7] M IP, WB Mm Hu ab1410814-3-3 beta P IHC-P, IP, WB Rb Hu, Mm, Rt ab1526014-3-3 beta P WB Rb Hu, Mm, Rt ab1673014-3-3 beta [60C10] M ELISA, WB Mm Hu, Mm, Rt ab1685914-3-3 beta, prediluted P IHC-P Rb Hu, Mm, Rt ab1526214-3-3 beta + epsilon + zeta [3C8] M WB Mm Hu, Mm, Rt ab1234614-3-3 beta + zeta P WB Rb Ma, Dm ab1411314-3-3 beta + zeta [22-IID8B] M ICC, IHC, WB Mm Hu, Mm, Rt, B, D, Mk, Pi, Rb, Sh ab1234714-3-3 eta + gamma + zeta P ELISA, WB Rb Many ab1412414-3-3 gamma (acetyl V1) [KC21] M WB Mm Hu ab1700514-3-3 gamma [3F8] M IP, WB Mm Hu ab1411714-3-3 sigma [1N6] M IF, IHC-P, IP, WB Mm Hu ab1412314-3-3 tau [3B9] M IP, WB Mm Hu, Mm, Rt, B ab1043914-3-3 Binding Motif (phospho S) P ELISA, IHC-P, IP, WB Rb Many ab1412714-3-3 Binding Motif (phospho S) [3i2] M ELISA, IP, WB Mm Many ab14126Agrin [AGR 131] M ICC, IHC, IRMA, WB Mm Mm, Rt ab12362Agrin [Agr 247] M ICC, IF Mm Mm, Rt ab12364Agrin [Agr 86] M FACS, ICC, WB Mm Mm, Rt ab12361AKT P WB Ch Hu, Mm, Rt, X , B ab13990AKT P ICC, IP, WB Rb Hu, Mm, Rt ab6076

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AKT (unmodified S473) [BDI111] M ELISA, WB Mm Hu ab19442AKT (phospho S473) P IF Rb Hu, Mm ab20488AKT (phospho T308) P IF, WB Rb Hu ab8933AKT (phospho T34) P ELISA, WB Rb Hu, Mm, Rt, Rb ab23509AKT peptide BL ab9041CrkL P WB Gt Hu, Mm ab10907CSN7b P WB Rb Hu ab11895Ctip1 [14B5] M ICC, IF, IHC, IP, WB Mm Hu, Mm, Fi ab19487Ctip1 [15E3AC11] M ICC, IF, IHC, IP, WB Mm Hu, Mm ab18688Ctip1 [18B12DE6] M ICC, IF, IHC, IP, WB Mm Hu, Mm ab19489

AKT (phospho T308) antibody (ab4796)

IHC - ab4796 at a dilution of 1/100 (2 overnight incubations at room temperature; ABC amplification). Cytoplasmic andnuclear staining observed in rat spinal cord motorneurons (ventral horn) 30µm saggital spinal cord sections.

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC Rb Hu, Mm, Rt www.abcam.com/ab4796

AKT (phospho S473) antibody (ab4802)

IF - detection of AKT phospho S473 at a dilution of 1/200 (24h incubation at RT). Perinuclear staining observed inventral horn neurons of rat spinal cord (30µm saggital sections).

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-P, IHC-Fr, IF, ICC, IHC Rb Hu, Mm, Rt www.abcam.com/ab4802

AKT (phospho S473) antibody (ab8932)

IF - AKT is phosphorylated on Ser 473 in breast tumor. The staining is much stronger than the weak basal level ofphosphorylation in normal breast. The staining is cytoplasmic, as in normal tissue. The phospho Ser 473 AKTantibody (ab8932) is used with no pretreatment at a dilution of 1:100.

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-P, IF Rb Hu www.abcam.com/ab8932

AKT antibody (ab8805)

IF - ab8805 used at 1/80 on cultured neonatal rat cardiomyocytes that express a nuclear-targeted AKT construct. AKTstaining (green) and actin filaments (red).

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-P, IF, ICC, IP, Rb Hu www.abcam.com/ab8805

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Dab1 P IP, WB Gt Mm ab16674Dab1 P IF, IHC-Fr, IP, WB Rb Mm ab7522Dab1 (phospho S491) P WB Rb Hu ab5776Dab1 (phospho Y198) P WB Rb Hu ab5673Dab1 (phospho Y220) P WB Rb Hu ab5677Dispatched P Fast track Gt Fast track ab10142ELAVL2 + ELAVL3 + ELAVL4 [16A11] M IHC-Fr, WB Mm Hu, Mm, Rt, Fi ab14370EMX2 P IHC, WB Rb Hu, Mm, Rt ab11849Fetuin A P ELISA, WB Rb Hu ab13980FMR1 (Drosophila) [6A15] M ELISA, IF, IP, WB Mm Dm.

Does not react with Hu ab10299FOXG1 P WB Gt Hu ab3394 Pep. avail.FOXG1 P IHC-P, WB Rb Hu ab23470GCNF P IHC-P Rb Hu ab13007Gemin 1 [2B1] M ELISA, IF, IP, WB Mm Hu, Mm, X ab5831Gemin 2 [2E17] M ELISA, IF, IP, WB Mm Hu, X ab6084Gemin 3 [12H12] M ELISA, IF, IP, WB Mm Hu ab10305Gli1 P IHC-P, IP, WB Rb Hu, Mm ab7523Gli2 P Fast track Rb Fast track ab7181Gli2 P Fast track Rb Fast track ab7195Gli3 P Fast track Rb Fast track ab6050LIM Kinase P WB Rb Mm ab10561LIS1 P IP, WB Rb Hu ab2607LIS1 P WB Gt Mm ab2656 Pep. avail.MAP4K6 P IHC-P Rb Hu ab13140NCAM [123C3] M IF, IHC-F, IHC-Fr, IHC-P Mm Rt, Hu ab8077NCAM [B332 (123A8)] M FACS, IHC-Fr, WB Mm Hu ab8596NCAM [ERIC-1] M ELISA, IHC-Fr, IP, RIA, WB Mm Hu ab6123NCAM [H28-123] M AP, IHC-Fr, IM, WB Rt Mm ab19782NCAM [MEM-188] M FACS, IP, WB Mm Hu ab8233NCAM [RNL-1] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt ab9018Neuro D4 P Fast track Gt Fast track ab3940 Pep. avail.Neuroglobin P ELISA, WB Ch Hu ab14010NR2E3 P IHC-P Rb Hu ab13081NRG1 P WB Rb Hu, Mm ab2994NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369PAX2 P IHC-P, WB Rb Hu ab23799PAX6 P WB Rb Rt, Sh ab5790 Pep. avail.PHOX2A P IHC, IP, WB Rb Hu, Mm, Rt ab12046PHOX2A P Fast track Gt Fast track ab4511PHOX2B P IHC, IP, WB Rb Rt, Hu, Mm ab12047PROX1 P ELISA, IHC-Fr Rb Hu, Mm, Rt, Ch, Fi ab11941RAI3 P ICC, IHC-P Rb Hu ab13274ROR alpha P WB Rb Hu ab16367ROR alpha P IHC-P Rb Hu ab13109ROR beta P IHC-P Rb Hu ab13113SCP1 P IF Rb Hu, Mm ab15087SCP1 P IF, IHC-Fr, IHC-P Rb Mm ab15090

Ctip2 antibody [25B6] (ab18465)

IHC - using ab18465 on adult mouse hippocampus.

Clonality Applications tested Host Species reactivity DatasheetM WB, IP, IHC-P, IHC-Fr, IF, Rt Hu, Mm, Fi www.abcam.com/ab18465

ICC, IHC, ChIP

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SIRP alpha P IHC-P, WB Rb Hu, Mk.Does not react with Mm ab2971

SIRP alpha P WB Rb Hu, Rt, Mm ab8120 Pep. avail.SIRP [MRC OX-41 ] M FACS, IHC-Fr, IHC-P, WB Mm Rt ab9295Slit1 P WB Rb Mm, Rt ab10984Slit2 P Fast track Rb Fast track ab7665SOX2 P IF Rb Hu, Mm ab15830 Pep. avail.SOX6 P IHC, IP, WB Rb Hu, Mm, Rt ab12054SOX8 P IHC, IP, WB Rb Hu, Mm, Rt ab12055SOX13 P IHC, IP, WB Rb Hu, Mm, Rt ab12060SOX14 P IHC, IP, WB Rb Hu, Mm, Rt ab12061SOX22 P IHC, IP, WB Rb Hu, Mm, Rt ab12062Sprouty 2 P ELISA, IP, WB Rb Hu ab1043SynCAM P WB Rb Rt ab3910Tenascin C P WB Ch Hu ab16290Tenascin C [MTn-12] M AF, Dot, ELISA, IF, IHC, IHC-F,

IHC-Fr, IP, WB Rt Mm ab6346TRAF6BP P WB Rb Hu ab22049VCAM1 [14C3] M IF, IHC-P Mm Hu, Mm ab11514VCAM1 [1G11B1] M FACS, IA, ICC, IHC-Fr, IP Mm Hu ab7219VCAM1 [M/K-2] M FuncS, IF, IHC Rt Mm ab19569VCAM1 [MVCAMA(429)] M FACS Rt Mm ab23807Zic1 P IHC, IP, WB Rb Hu, Mm, Rt ab12070Zic2 P IHC, WB Rb Hu, Mm, Rt ab12072

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Neuroscience > Neurodegeneration > Alzheimer’s > Amyloid


ADAM10 P WB Rb Hu ab10956AMPK alpha 1 P IP, WB Rb B, Hu, Rt ab3759Amyloid A Component [mcl], prediluted M IHC-Fr, IHC-P Mm Hu ab7505Amyloid beta Precursor Protein P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab17468Amyloid beta Precursor Protein P ICC, IP, WB Rb Hu, Mk ab12268Amyloid beta Precursor Protein P ICC, IHC-P, IP, WB Rb Hu, Mk ab12269Amyloid Precursor Protein P ELISA, IHC-P, WB Gt Hu ab17325Amyloid Precursor Protein P IHC-P, IP, WB Rb Hu, Mk ab11117Amyloid Precursor Protein P WB Rb Hu, Mm ab2073Amyloid Precursor Protein P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab2084Amyloid Precursor Protein (phospho T668) P ICC, IHC-P, IP, WB Rb Hu, Mm, Rt ab17417Amyloid Precursor Protein [3E9] M IHC-P, WB Mm Hu ab12338Amyloid Precursor Protein Frameshift Mutant P ELISA, IHC-P, WB Gt Hu ab2085Amyloid Precursor Protein peptide BL ab7874APH1a P IF, IP, WB Rb Hu, Mm ab19390APH1a P WB Rb Rt, Hu, Mm ab12104 Pep. avail.beta Amyloid 1-40 P ELISA, IHC-Fr, RIA Rb Hu ab16218beta Amyloid 1-40 P IP, WB Rb Hu, Mm ab16622beta Amyloid 1-40 P ELISA, IHC-P, WB Rb Hu ab10147beta Amyloid 1-40 [3H2] M ELISA, IF, IHC-P, WB Mm Hu ab17331beta Amyloid 1-40 [BAM-10] M IHC-Fr, IHC-P Mm Hu ab7501beta Amyloid 1-40 [BDI350] M ELISA, ICC, WB Mm Hu ab20068beta Amyloid 1-42 P ELISA, IHC-P, WB Rb Hu ab10148beta Amyloid 17-42 P ELISA Rb Hu ab17327beta Amyloid P ELISA, IHC, WB Ch Hu ab17472beta Amyloid P IHC Rb Ro ab14220beta Amyloid P ELISA, WB Rb Hu, Mm ab2539beta Amyloid [2C8] M IHC, WB Mm Hu, Mm ab12349beta Amyloid [4G8] M ELISA, IHC-P, IP, WB Mm Hu ab1910beta Amyloid peptide BL ab3698CLACP P WB Rb Hu, Mm ab16763Factor VIII heavy chain [RFFVIIIC/8] M ELISA, WB Mm Hu, Pi.

Does not react with Mm, Rt or D ab6345

beta Amyloid antibody [6E10] (ab10146)

IHC-P- using ab1910 [clone 4G8] and ab10146 [clone 6E10]. Stains paraffin - embedded cerebellum.

Clonality Applications tested Host Species reactivity DatasheetM WB, IP, ELISA, IHC-P Mm Hu www.abcam.com/ab10146

Amyloid Precursor Protein antibody [MABDE2B4] (ab12266)

IHC-P- ab12266 detecting APP in Alzheimer’s disease human brain. Plaques are observed in the neuropil. Amyloiddeposits stained by DE2B4 (ab12266) can be seen around some blood vessels.

Clonality Applications tested Host Species reactivity DatasheetM WB, IP, IHC-P, ICC, Mm Hu, C, Mk, Pi www.abcam.com/ab12266

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FE65 P WB Rb Mm ab5668 Pep. avail.FE65 [4H324] M ICC, IP, WB Mm Hu, Mm, Rt ab17469Humanin P WB Rb Hu ab16758Mint1 P WB Rb Rt ab3448 Pep. avail.Mint3 P WB Rb Rt, Mm ab3450 Pep. avail.PEN2 P IP, WB Rb Hu ab16555RAGE P ELISA, IHC-P Gt Hu ab20558RAGE P IHC-Fr, WB Rb Rt, Mm ab3611SHC P IP, WB Rb Hu, Mm, Rt ab15039SHC (phospho Y239 + Y240) P WB Rb Hu ab4890 Pep. avail.SHC (phospho Y317) P WB Rb Hu, Mm ab15040

Neuroscience > Neurodegeneration > Alzheimer’s > Related targets


Acrolein P ELISA Rb Acrolein ab15138Caspase 6 (active) P WB Rb Mm, Rt, Hu ab2326ADAM17 P Dot, WB Rb Hu, Rt, Mm ab2051 Pep. avail.ADAM17 P WB Rb Hu, Rt, Pi ab11547ADAM17 P WB Gt Hu ab13535 Pep. avail.Als2 P Fast track Gt Fast track ab4155 Pep. avail.ALS2CR1 P WB Gt Hu ab4123 Pep. avail.ALS2CR2 P WB Gt Hu ab4122 Pep. avail.ALS2CR2 P WB Rb Hu, Mm, Rt ab17205ALS2CR3 P Fast track Gt Fast track ab4200 Pep. avail.Apolipoprotein E P ELISA, IHC-P, WB Gt Hu ab7620Apolipoprotein E P ID, IE, RID, WB Rb Mm ab20874Apolipoprotein E [E6D7] M ELISA, IHC-Fr, IHC-P, IP, WB Mm Hu ab1907Calpastatin P ELISA, WB Ch Hu ab16423Calpastatin [2G11D6] M ICC, WB Mm Hu, Rt, C, Pi ab3515Caspase 6 P WB Rb Hu, Mm, Rt ab2552Caspase 6 P ELISA, WB Rb Hu, Mm, Rt ab18520Caspase 6 P WB Rb Hu ab19920Caspase 6 [3F51] M IHC-P, IP, WB Mm Hu ab17866Caspase 6 [3F52] M IHC-P, IP, WB Mm Hu ab17822Caspase 6 [6CSP01 (7165)] M IP Mm Hu ab3245Caspase 6 [6CSP03] M WB Mm Hu ab4062CCKAR P WB Rb Hu ab14441Drebrin P WB Rb Rt, Hu ab11068Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350ERAB P ELISA, IF, IHC-P, WB Rb Hu ab17298ERAB [5F3] M Dot, ELISA, IF, IHC-Fr, IHC-P, WB Mm Hu ab10260Fyn P ELISA, WB Gt Hu ab802 Pep. avail.

ApoE antibody [D6E10] (ab1906)

IHC-P - ab1906 reactivity with Apo E of vascular amyloid beta in a Down Syndrome tissue section.

Clonality Applications tested Host Species reactivity DatasheetM WB, IHC-P, IHC-Fr Mm Hu www.abcam.com/ab1906

Neurodegeneration


Neuroscience > Neurodegeneration > Alzheimer’s > Amyloid


20

Fyn P WB Rb Hu, Mm ab13955Fyn [1S] M IHC-P Mm Hu, Rt, Mm ab3116Fyn [FYN-01] M ICC, IP, WB Mm Hu, Mm ab1881PHF2 P WB Rb Hu ab1138Synuclein (alpha) P H, ICC, IHC, IHC-Fr, IHC-P, WB Sh Hu ab6162Synuclein (alpha) P IHC-P Rb Hu, Rt ab15530Synuclein (alpha) P ICC, WB Gp Hu, Mm, Rt, B ab16784Synuclein (alpha) (phospho Y125) P Fast track Rb Fast track ab10789 Pep. avail.Synuclein (alpha) [4B12] M Dot, ELISA, IHC-Fr, IHC-P, WB Mm Hu.

Does not react with Mm or Rt ab1904Synuclein (alpha) [4D6] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt ab1903Synuclein (beta) P ICC, IHC-Fr, IHC-P, WB Rb Hu ab6165Synuclein (beta) P IHC-Fr, IHC-P Sh Hu ab6163Synuclein (beta) P WB Ch Hu ab16420Synuclein (beta) P IHC-P Rb Hu, Rt ab15532Synuclein (gamma) P IHC, IHC-Fr, IHC-P, WB Rb Hu, Rt ab6169Synuclein (gamma) P IHC-Fr, IHC-P, WB Rb Hu, Rt ab6176Synuclein (gamma) P IHC-P, WB Rb Hu, Rt ab15534Ubiquitin P ELISA, IF, IHC-F, IHC-Fr,

IHC-P, IP, WB Rb Hu, Rt, H, Mm, B ab7780Ubiquitin P ELISA, WB Rb Y ab14372Ubiquitin P WB Ch Hu ab19310Ubiquitin [10C2-2] M AP, ELISA, IHC-Fr, IHC-P, WB Mm All ubiquitinated proteins ab411Ubiquitin [1B4-UB] M ELISA, FACS, IHC-Fr, IHC-P, WB Mm Hu, B ab122

Neuroscience > Neurodegeneration > Alzheimer’s > Proteases


BACE1 P IHC-P Gt Hu ab11028BACE1 P WB Rb Hu, Rt, Mm ab2077 Pep. avail.BACE2 P WB Rb Hu, Rt, Mm ab8025 Pep. avail.BACE peptide BL ab5857Cathepsin H [1D10] M IHC-Fr, IHC-P, WB Mm Hu ab7432Nicastrin P WB Rb Hu, Mm ab3444 Pep. avail.Presenilin 1 P ELISA, IHC-Fr, IHC-P, WB Gt Hu ab7297Presenilin 1, prediluted P IHC-P Rb Hu ab15508Presenilin 1 [APS 11] M ELISA, IF, IHC-P, WB Mm Hu, Mm, Rt ab15456Presenilin 1 [APS 18] M ELISA, IF, WB Mm Hu, Mm, Rt ab15458Presenilin 2 P ELISA, IHC, WB Ch Hu, Mm, Rt ab18041Presenilin 2 P ELISA, IHC-Fr, IHC-P Gt Hu ab11148Presenilin 2 [198C67921] M WB Mm Hu ab13926Presenilin 2 [APS 26] M ELISA, IF, WB Mm Hu, Mm, Rt ab15549

Ubiquitin antibody (ab6309)

IF - P23H mutant cells immunostained with ab6309 (1/500 dilution) showing co-localization of mutant aggregatedprotein with ubiquitin.

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-Fr, IA, IF Rb C, Sc www.abcam.com/ab6309

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Neuroscience > Neurodegeneration > Alzheimer’s > Related targets


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Neuroscience > Neurodegeneration > Alzheimer’s > Tangles & Tau


14-3-3 tau [3B9] M IP, WB Mm Hu, Mm, Rt, B ab10439Casein Kinase 1 delta P ELISA, IHC-P Gt Hu ab10877ERK1 P IHC-P, IP, WB Rb Hu, Rt, Mm ab7947 Pep. avail.ERK1 P ELISA, IP, WB Rb Hu, Rb, Rt, Mm, C, Pi ab9363ERK1 (HRP) P ELISA, IP, WB Rb Hu, B, Mm, Pi, Rb, Rt ab9364ERK1 + ERK2 (phospho T185 + T202) P WB Rb Hu ab4819 Pep. avail.ERK1 + ERK2 (phospho T185 + T202) (FITC) P FACS, IHC-Fr, WB Rb Hu ab4820ERK1 + ERK2 (phospho T202/183 + Y204/185) P WB Rb Hu, Mm, Rt ab16869ERK1 + ERK2 (phospho T202 + Y204) P IF Rb Hu, Mm, Rt ab20490ERK1 + ERK2 P IP Sh Hu, Rt ab16573ERK1 + ERK2 P WB Rb Hu, Mm, Rt, B ab17942GSK3 (alpha + beta) (phospho Y216 + Y279) P Dot, ELISA, WB Rb Hu, Mm, Rt ab4797GSK3 (alpha + beta) P IHC-P, WB Rb Hu, Mm, Rt, Ch, Mk ab15314GSK3 (alpha + beta) [1H8] M ELISA, WB Mm Hu, Mm, Rt, Am ab17059Neurofibrillary Tangle P IHC-Fr, IHC-P, WB Rb Hu ab2076Neurofibrillary Tangle P ELISA, IHC-P, WB Rb Hu ab11093PEN2 P IP, WB Rb Hu ab16555Pin1 P WB Rb Hu ab12107 Pep. avail.Prealbumin P DID, Ie, RID, RIe Sh Hu, B, F, Ch, D, Gt, Gp, H,

Mm, Pi, Rb, Rt ab9015Prealbumin P IHC, RID Rb Hu, B, F, D, H ab16006Prealbumin P ELISA Gt Hu ab15007Tau (phospho S199) P WB Rb Hu ab4749 Pep. avail.Tau (phospho S199 + S202) P WB Rb Hu ab4864Tau (phospho S202) P IHC-Fr, WB Rb Hu ab4839 Pep. avail.Tau (phospho S208) P WB Rb Hu ab10892 Pep. avail.Tau (phospho S214) P WB Rb Hu ab4846 Pep. avail.Tau (phospho S214) P Fast track Rb Fast track ab10891 Pep. avail.Tau (phospho S356) P WB Rb Hu ab4857 Pep. avail.Tau (phospho S262) P WB Rb Hu ab4856 Pep. avail.Tau (phospho S396) P WB Rb Hu ab4858 Pep. avail.Tau (phospho S400) P WB Rb Hu ab4859 Pep. avail.Tau (phospho S404) P WB Rb Hu ab4860 Pep. avail.Tau (phospho S409) P WB Rb Hu ab4861 Pep. avail.Tau (phospho S422) P WB Rb Hu ab4862 Pep. avail.Tau (phospho T181) P WB Rb Hu ab4748 Pep. avail.Tau (phospho T212) P WB Rb Hu ab4842 Pep. avail.Tau (phospho T217) P WB Rb Hu ab4851 Pep. avail.Tau (phospho T231) P WB Rb Hu ab4855 Pep. avail.Tau P IHC-P, WB Gt Hu ab11107Tau P IHC-F, WB Rb Hu ab22690Tau P IF, IHC-Fr, IHC-P Rb Hu ab8763Tau [T1] M WB Mm Hu, Mm, Rt, B ab12354Tau [Tau] M IHC-Fr, IHC-P Mm Ch, B, Hu, Mk.

Does not react with Mm or Rt ab8739Tau [TAU-5] M IHC-Fr, IHC-P, WB Mm Hu ab3931

Tau antibody (ab4841)

IF - Transgenic C. elegans first larval stage nematode expressing human wild type Tau GFP (green) and stained withab4841 (red). Neurons of the head, dorsa and ventral cords and tail region display phosphorylated Tau.

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-Fr, IHC (no paRtffin) Rb Hu www.abcam.com/ab4841

whole animals

Neurodegeneration


22

Ubiquitin P IA, IF, IHC-Fr, WB Rb C, Sc ab6309Ubiquitin P ELISA, IF, IHC-F, IHC-Fr,

IHC-P, IP, WB Rb Hu, Rt, H, Mm, B ab7780Ubiquitin P ELISA, WB Rb Y ab14372Ubiquitin P WB Ch Hu ab19310Ubiquitin [10C2-2] M AP, ELISA, IHC-Fr, IHC-P, WB Mm All ubiquitinated proteins ab411Ubiquitin [1B4-UB] M ELISA, FACS, IHC-Fr, IHC-P, WB Mm Hu, B ab122

Neuroscience > Neurodegeneration > Huntington’s


HAP40 P IP, WB Rb Mm ab1139HAP40 P ELISA, WB Rb Hu ab1400HIPPI P WB Rb Hu ab5205Huntingtin [1A771] M WB Mm Many ab13583Huntingtin [HDA3E10] M IF, IHC-Fr, IP, WB Mm Hu, Rt, Mm ab7668Huntingtin Interacting Protein HIP1 [4B10] M WB Mm Hu ab5402Huntingtin Interacting Protein HIP1[4B107], prediluted M IHC-P Mm Hu.

Does not react with Mm ab11841Huntingtin Interacting Protein 2 P ELISA, WB Gt Hu ab21728

Neuroscience > Neurodegeneration > Parkinson’s > Related targets


ALDH1A1 P WB Gt Hu, Mm.Does not react with Rt ab9883 Pep. avail.

APH1a P WB Rb Rt, Hu, Mm ab12104 Pep. avail.APH1a P IF, IP, WB Rb Hu, Mm ab19390CCKAR P WB Rb Hu ab14441DORFIN P Fast track Gt Fast track ab3997 Pep. avail.DORFIN P Fast track Rb Fast track ab10889

ALDH1A1 antibody (ab24343)

IF - staining of ALDH1A1-containing neurons in adult mouse brain sections using ab24343.

Clonality Applications tested Host Species reactivity DatasheetP IHC-Fr, IHC-FrFl Rb Mm www.abcam.com/ab24343

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Neuroscience > Neurodegeneration > Alzheimer’s > Tangles & Tau


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PGP95 P WB Gt Hu ab11145PGP95 P IHC-Fr Gp Hu, Mm, Rt ab10410PGP95 P IHC-P Rb Hu ab15503PGP95 [13C4 / I3C4] M IF, IHC-F, IHC-Fr, IHC-P Mm Hu, Rt, Gp ab8189PGP95 [31A3] M ELISA, IHC-P, WB Mm Hu, Rt ab20559PINK1 P IHC-P, WB Rb Hu, Mm, Rt ab23707SIAH1 P WB Gt Hu ab2237 Pep. avail.Synphilin 1 P IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt ab6179Synphilin 1 P ELISA, IHC-P, WB Gt Hu ab11106

Neuroscience > Neurodegeneration > Parkinson’s > Parkin / PARK


PARK7 P WB Gt Hu, Mm ab4150 Pep. avail.PARK7 [malphaDJ-1/E219] M ICC, WB Mm Hu, Fi.

Does not react with Mm ab11251Parkin P IHC-Fr, WB Rb Hu, Rt ab5667 Pep. avail.Parkin P Dot, ELISA, IHC-Fr, IHC-P, WB Rb Hu, Rt ab6177Parkin P ELISA, IHC-P, WB Gt Hu ab7296

Neuroscience > Neurodegeneration > Parkinson’s > Synuclein


DORFIN P Fast track Rb Fast track ab10889DORFIN P Fast track Gt Fast track ab3997 Pep. avail.HMGB1 [4C3] M WB Mm Hu, Mm, Rt, C, Pi ab11354HMGB1 [HAP465] M ELISA, WB Mm Hu, Mm, Rt, Ch, D ab12029Synphilin 1 P ELISA, IHC-P, WB Gt Hu ab11106Synphilin 1 P IHC-Fr, IHC-P, WB Rb Hu ab6179Synuclein (alpha) P ICC, WB Gp Hu, Mm, Rt, B ab16784

Parkin antibody (ab15494)

IHC - ab15494 staining Parkin in Alzheimer’s brain (FFPE-sections).

Clonality Applications tested Host Species reactivity DatasheetP IHC-P Rb Hu, Mm, Rt www.abcam.com/ab15494

PGP9.5 antibody (ab10404)

IF - ab10404 at a dilution of 1/1000, detecting PGP9.5 in cortical neurons in rat brain tissue sections prepared for freefloating IHC (4% PFA fixed, 30µm thick coronal sections).

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-Fr, IF Rb Hu, Mm, Rt, Pi www.abcam.com/ab10404

Neurodegeneration


Neuroscience > Neurodegeneration > Parkinson’s > Related targets


24

Synuclein (alpha) P IHC-P Rb Hu, Rt ab15530Synuclein (alpha) P H, ICC, IHC, IHC-Fr, IHC-P, WB Sh Hu ab6162Synuclein (alpha) (phospho Y125) P Fast track Rb Fast track ab10789 Pep. avail.Synuclein (alpha) [4B12] M Dot, ELISA, IHC-Fr, IHC-P, WB Mm Hu.

Does not react with Mm or Rt ab1904Synuclein (beta) P IHC-P Rb Hu, Rt ab15532Synuclein (beta) P WB Ch Hu ab16420Synuclein (beta) P IHC-Fr, IHC-P Sh Hu ab6163Synuclein (beta) P ICC, IHC-Fr, IHC-P, WB Rb Hu ab6165Synuclein (gamma) P IHC, IHC-Fr, IHC-P, WB Rb Hu, Rt ab6169Synuclein (pan) P IHC-P, WB Rb Hu, Rt ab15534Synuclein (pan) P IHC-Fr, IHC-P, WB Rb Hu, Rt ab6176VCP [5] M IHC-Fr, IHC-P, IP, WB Mm Hu, Mm, Rt ab11433

Neuroscience > Neurodegeneration > Prions


67kDa Laminin Receptor P ICC, IF, IHC-P, IP, WB Rb Hu, Mm ab71167kDa Laminin Receptor [MLuC5] M FACS, ICC, IF, IHC-P Mm Hu, Rt ab3099Fyn P WB Rb Hu, Mm ab13955Fyn P ELISA, WB Gt Hu ab802 Pep. avail.Fyn [1S] M IHC-P Mm Hu, Rt, Mm ab3116Fyn [FYN-01] M ICC, IP, WB Mm Hu, Mm ab1881Prion protease resistant protein 27-30 P ELISA, IHC-P, WB Gt Hu ab7303Prion protein PrP P ELISA, WB Mm Hu, B ab22366Prion protein PrP P ELISA, WB Rb Hu, Mm, B ab703Prion protein PrP P Dot, ELISA, IHC, WB Gt Hu, B, Sh, Ha ab6664Prion protein PrP [1E5 / G6] M ELISA, WB Mm Hu, B ab2879Prion protein PrP [1E5 / G6] M ELISA, WB Mm Hu, B ab3408Prion protein PrP [3B8 / D5] M ELISA, WB Mm C, Sh ab2881Prion protein PrP [7D9] M ELISA, IHC, IHC-F, IHC-P, WB Mm Mm, B, De, El, Ha, Hu, Rt ab14219Prion protein PrP [WD3C7] M ELISA, IP, WB Mm B ab13643

alpha Synuclein [4D6] (ab1903)

IHC-P - ab1903 stains Lewy Bodies in a substantia nigra neuron.

Clonality Applications tested Host Species reactivity DatasheetM ELISA, WB, IHC-P, IHC-Fr Mm Hu, Mm, Rt www.abcam.com/ab1903

HMGB1 (ab11972)

ICC- labeling of HMGB1 (red) in HeLa cells using ab11972 (1/50). DNA is stained with DAPI (blue).

Clonality Applications tested Host Species reactivity DatasheetP IF, WB Rb Hu www.abcam.com/ab11972

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Neuroscience > Neurodegeneration > Parkinson’s > Synuclein


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ProteinsPrion protein ab1391Prion protein ab753

Neuroscience > Neurodegeneration > Related targets


5HT2A Receptor P IHC-FrFl, WB Rb Rt ab16028 Pep. avail.Als2 P Fast track Gt Fast track ab4155 Pep. avail.ALS2CR1 P WB Gt Hu ab4123 Pep. avail.ALS2CR2 P WB Gt Hu ab4122 Pep. avail.ALS2CR2 P WB Rb Hu, Mm, Rt ab17205ALS2CR3 P Fast track Gt Fast track ab4200 Pep. avail.Calpain 6 P ELISA, IHC-Fr, IP, WB Rb Hu, Mm, Rt ab16350CRMP5 [CR-1] M IHC-Fr, WB Rt Hu, Rt, B ab19378CRMP5 [CR-3] M IHC-Fr, WB Rt Hu, Mm, Rt, B ab19352Neuroserpin P IHC, WB Rb Hu ab16171Ubiquitin P ELISA, WB Rb Y ab14372Ubiquitin P ELISA, IF, IHC-F, IHC-Fr, IHC-P, IP, WB Rb Hu, Rt, H, Mm, B ab7780Ubiquitin P IA, IF, IHC-Fr, WB Rb C, Sc ab6309Ubiquitin P WB Ch Hu ab19310Ubiquitin [10C2-2] M AP, ELISA, IHC-Fr, IHC-P, WB Mm All ubiquitinated proteins ab411Ubiquitin [1B4-UB] M ELISA, FACS, IHC-Fr, IHC-P, WB Mm Hu, B ab122

HMG14 antibody (ab5212)

ICC - Labeling of HMGN1 (green; ab5212, 1/500) and monoclonal mouse anti-beta-tubulin-II (red) in fixed primarycultures of rat (e18) hippocampal neurons revealed exclusive nuclear staining corresponding to the known nuclearlocalization of HMG14.

Clonality Applications tested Host Species reactivity DatasheetP IF, WB Rb Hu, Mm, Rt www.abcam.com/ab5212

67kD Laminin Receptor antibody [SPM278], prediluted (ab18099)

IHC-P - Human breast carcinoma stained with ab18099.

Clonality Applications tested Host Species reactivity DatasheetM IHC-P Mm Hu www.abcam.com/ab18099

Neurodegeneration


Neuroscience > Neurodegeneration > Prions


26

Neuroscience > Neuroendocrinology > GH Regulation


ENSA P WB Ch Hu, Mm, Rt, B ab14297GHRHR P IHC-P Rb Hu ab13342Somatostatin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7840Somatostatin 14 P Dot, WB Rb Rt ab8903Somatostatin 14, prediluted P IHC-P Rb Hu ab15522Somatostatin 28 P Dot, RIA, WB Rb Rt ab8904Somatostatin Receptor P IHC-Fr, IHC-P Rb Rt, Mm ab2366Somatostatin Receptor 1 P FACS, WB Rb Hu, Rt ab5827Somatostatin Receptor 1 P IP, WB Rb Hu, Mm, Rt ab479Somatostatin Receptor 2 P IF, WB Rb Hu, Mm, Rt ab9550Somatostatin Receptor 2 P IF, IHC-P Rb Hu, Rt ab13120Somatostatin Receptor 4 P ELISA, WB Rb Rt ab18796

ProteinsSomatostatin 28 protein ab9606Somatostatin protein ab9607

Neuroscience > Neuroendocrinology > Gonadotrophic axis


Estradiol P RIA Sh Many ab22441Estradiol [6E1] M RIA Mm Hu ab20257Estradiol [B42] M ELISA, IHC-Fr Mm Ma ab8743Estrogen Receptor alpha M IHC-Fr, WB Rb Hu, B ab8617Estrogen Receptor alpha P WB Ch Hu ab14020Estrogen Receptor alpha P IP, WB Mm Hu ab283Estrogen Receptor alpha P ELISA, IHC-P, WB Rb Hu ab5700Estrogen Receptor alpha [1D5] M IHC-Fr, IHC-P Mm Hu, Rt ab858Estrogen Receptor alpha [B301] M IHC-F, IHC-Fr, IHC-P, WB Mm Hu ab8857Estrogen Receptor alpha [B401 (AER303)] M ELISA, IHC-Fr, IP, WB Mm Hu, B ab7820Estrogen Receptor alpha [EVG F9] M IHC-P, IP, WB Mm Hu, Rb ab19349Estrogen Receptor alpha [h-151] M GSA, IHC, IP, WB Mm Hu, Mm, Rt, B ab13538

Estrogen Receptor alpha antibody [33] (ab2746)

ICC - mouse 4TI cells stained with ab2746 (1:100). Blue is DAPI stained nuclei, red is Estrogen Receptor alpha.

Clonality Applications tested Host Species reactivity DatasheetM WB, IP, IHC-P, GSA, FACS, IF Mm Hu, Rt, Mm, C www.abcam.com/ab2746

Somatostatin 28 antibody (ab22682)

EM - ab22682 (1/2000) in guinea pig pancreas D cells. On grid immunogold procedure and 10nm gold particles.

Clonality Applications tested Host Species reactivity DatasheetP EM, IF, IHC-Fr, IHC-P Rb Hu, Mm, Rt, Ch, Cow, Dog, www.abcam.com/ab22682

Cat, Guinea pig, Pig and Sheep

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Estrogen Receptor alpha (phospho S167) P ELISA, IP, WB Rb Hu ab5701Estrogen Receptor beta P WB Ch Hu, Mm, Rt ab14021Estrogen Receptor beta P GSA, IHC-Fr, IHC-P, WB Rb Hu, Rt, Mm, Mk ab3577 Pep. avail.Estrogen Receptor beta P IHC, WB Rb Rt ab5784Estrogen Receptor beta P IHC-Fr, IHC-P, WB Rb Hu, Rt ab5786Estrogen Receptor beta [14C8] M FACS, IHC, IHC-P, WB Mm Hu ab288Estrogen Receptor beta [988] M IP, WB Mm Hu, Mm, Rt ab16813Progesterone Receptor P IHC-P Rb Hu ab15509Progesterone Receptor [Alpha PR6] M ICC, IHC-Fr, IP, WB Mm Hu, Rb, Rt, Mm, B, Gp, Ch ab2765Progesterone Receptor [PGR-1A6] M IHC-Fr, IHC-P Mm Hu ab821Progesterone Receptor [PR-AT 414] M IHC-Fr, IP, WB Mm Hu, Rb, Rt, Mm, B ab2764 Pep. avail.Progesterone Receptor [SP2] M IHC-P, WB Rb Hu ab16661Progesterone Receptor (phospho S190) P Dot, WB Rb Hu ab13877Progesterone Receptor (phospho S190) [1154] M WB Mm Hu ab9895Progesterone Receptor (phospho S294) P Dot, WB Rb Hu ab13878Progesterone Receptor (phospho S294) [608] M WB Mm Hu ab9896

Neuroscience > Neuroendocrinology > HPA axis


ACTH P Dot, IHC-Fr Rb C, Rt ab8906ACTH P IHC-P Rb Hu ab10335ACTH P RIA Rb Hu, Rt, Mk ab11116ACTH P ICC, WB Ch Hu, Mm, Rt ab14064ACTH [56] M ELISA, ICC, IHC-P Mm Hu, Rt ab21003ACTH [A2H8] M ELISA, RIA Mm Hu ab11114ACTH [B427] M IF, IHC-F, IHC-Fr, IHC-P, ICC, WB Mm Hu ab8615CCKBR P IHC-P Rb Hu ab13173CCKBR P WB Rb Hu ab14439Corticoliberin [4H9] M ELISA Mm Hu ab10034Corticotropin Releasing Factor P AP, ICC, RIA Rb Hu, Rt ab8901Corticotropin Releasing Factor P IHC-Fr Rb Hu, Rt, Sh ab11133Corticotropin Releasing Factor P IHC, WB Ch Hu, Mm, Rt ab17475Dehydroepiandrosterone P RIA Rb Hu ab10999MC4 Receptor P ICC, IHC-Fr, IHC-P Rb Hu ab13034MC4 Receptor P IHC-P, WB Rb Hu, Mm, Rt ab24233Melatonin Receptor 1A P IHC-P Rb Hu ab13035NMUR1 P IHC-P Rb Hu ab13365NMUR2 P IHC-P Rb Hu ab13366

ProteinsACTH protein ab9586

Corticotropin Releasing Factor Receptor 2 antibody (ab12964)

IF - ab12964 (1/4000) detecting hypothalamic paraventricular CRF Receptor 2-containing neurons in rat brain.

Clonality Applications tested Host Species reactivity DatasheetP IF, IHC-P Rb Hu, Rt www.abcam.com/ab12964

Neuroendocrinology


Neuroscience > Neuroendocrinology > Gonadotrophic axis


28

Neuroscience > Neuroendocrinology > Obesity and Metabolism


Adiponectin P IP, WB Rb Mm.Does not react with Hu ab3455

Adiponectin P WB Rb Rt ab5596Adiponectin P ELISA, WB Rb Hu ab17895Adiponectin P ELISA, Neut, WB Gt Hu ab18065Adiponectin [19F1] M ELISA, IF, WB Mm Hu, Mm ab22554Adiponectin [NeNa] M ELISA, IP, WB Mm Hu.

Does not react with Mm ab16086Adiponectin Receptor 1 P WB Mm Mm, Rt ab21814Attractin P ELISA, WB Rb Mm ab14567Carboxypeptidase H P ICC, IHC-Fr, IP, WB Rb Hu, Mm, Rt, C, Mk ab11053CCKAR P WB Rb Hu ab14441Fatty Acid Synthase P IP, WB Rb Hu, Mm, Pi ab3844GALR2 P IHC-P Rb Hu ab13179Gastrin Releasing Peptide P IHC-Fr Rb Pi ab22623Ghrelin P ELISA Rb Hu ab14481Ghrelin P ELISA, WB Ch Hu, Mm, Rt ab15860Ghrelin P WB Rb Hu ab19314Ghrelin [Ghr2] M ELISA, WB Mm Hu ab13982Ghrelin [Ghr6] M ELISA, WB Mm Hu.

Does not react with Mm ab13984IL6 P ELISA, WB Ch Hu ab14038IL6 P WB Rb Pi ab11746IL6 P ELISA, Neut, WB Gt Rt ab9770IL6 P ELISA, WB Gt Rt ab17494IL6 P ELISA, Neut, WB Gt Hu ab17506IL6 [5E1] M ELISA, IHC-Fr, Inhib, WB Mm Mk ab8472IL6 [AS12] (FITC) M FACS Mm Hu.

Does not react with Mm ab16169IL6 [B-E8 ] M ELISA, IHC-Fr, Neut Mm Hu.

Does not react with Mm ab11449IL6 [MP5-20F3] M EIA, Inhib, Neut Rt Mm.

Does not react with Hu or Rt ab7375IL6 [MP5-32C11] M IHC-Fr, Neut Rt Mk.

Does not react with Hu or Rt ab7376

IL6 antibody (ab6672)

IHC - ab6672 staining in mouse collagen-induced arthritic tissue.

Clonality Applications tested Host Species reactivity DatasheetP ELISA, RIA, WB, IP, IHC Rb Hu, Mm, Rt www.abcam.com/ab6672

GLP1R antibody (ab13181)

IHC-P - ab13181 staining pancreatic formalin fixed paraffin embedded tissue.

Clonality Applications tested Host Species reactivity DatasheetP IHC-P, WB Rb Hu, Rt, Mm, Hamster www.abcam.com/ab13181

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IL6 [MQ2-13A5] (FITC) M FACS Rt Hu ab21513IL6 [MQ2-13A5] (PE) M FACS Rt Hu ab21514IL6 [MQ2-13A5] M ELISA, FACS, IHC-F, WB Rt Hu ab21515Insulin Receptor (phospho Y972) P WB Rb Hu, Mm ab5678Insulin Receptor P ELISA, WB Rb Hu ab5500Insulin Receptor [29B4] M IP Mm Hu, Mm, Rt ab16884LAR P WB Rb Hu ab10557LAR peptide BL ab22637Leptin P ICC, IHC-Fr, WB Rb Hu, Rt, Mm, Sh ab3583Leptin P ELISA, WB Ch Mm, Rt ab17531Leptin P ELISA, WB Rb Mm, Rt ab17629Leptin [9C10] (HRP) M ELISA Mm Hu ab6059Leptin [BDI142] M ELISA, ICC, IF, IHC-Fr Mm Hu ab20115Leptin [BDI170] M IF, IHC-Fr, WB Mm Hu ab20101Leptin Receptor P IHC-F, WB Rb Hu, Mm, Rt, D ab5593MC4 Receptor P ICC, IHC-Fr, IHC-P Rb Hu ab13034Melatonin Receptor 1A P IHC-P Rb Hu ab13035Melatonin Receptor 1B P IHC-P Rb Hu ab13358Melatonin Receptor 1B P IHC-P Rb Hu ab13357Neuropeptide EI NH2 P IHC-Fr Rb Rt ab11141Neuropeptide Y P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Gp, Rb ab6173Neuropeptide Y P Dot, IHC-Fr, RIA Rb Hu, Mm, Rt, F, B, Mk, Pi, Rb ab10980Neuropeptide Y P IHC-Fr Gp Rt ab10341NMUR1 P IHC-P Rb Hu ab13365NMUR2 P IHC-P Rb Hu ab13366Orexin A P ELISA, RIA Rb Hu, Mm, Rt ab10981Orexin A P ELISA, ICC, IHC-Fr Rb Hu ab16787Orexin B P ELISA, IHC, WB Rb Mm ab14970Orexin B P ELISA, RIA Rb Hu, Mm, Rt ab10982Prohormone Convertase 1 P WB Rb Mm ab3532 Pep. avail.Prohormone Convertase 1 P WB Rb Hu, Mk ab16020Prohormone Convertase 2 P WB Rb Mm ab3533 Pep. avail.Prohormone Convertase 2 P WB Rb Hu, Mk ab12275PTPIA2 / PTPRN P WB Rb Hu ab12154SLC6A4 [4A22] M FACS, IHC-P Mm Hu, Rt ab1125UCP1 P IHC-P, WB Rb Hu, Mm, Rt ab10983

ProteinsAdiponectin protein (His) ab13882IL6 protein ab9731IL6 protein ab9771IL6 protein ab9627Leptin protein ab646Leptin protein ab7651Leptin protein ab13987Leptin protein ab9827Leptin protein ab9750Neuropeptide Y protein ab9599

NPY5R antibody (ab13199)

IHC-P - ab13199 staining Hek-293 cells transiently transfected with recombinant NPY5R.

Clonality Applications tested Host Species reactivity DatasheetP ICC, IHC-P Rb Hu www.abcam.com/ab13199

Neuroendocrinology


Neuroscience > Neuroendocrinology > Obesity and Metabolism


30

Neuroscience > Neuroendocrinology > Oxytocin & Vasopressin


AVPR1B P IHC-P Rb Hu ab13147Oxytocin P IHC-P, RIA Rb Hu, Rt, Mm, Rb, Sh ab2078Oxytocin P IHC-Fr Rb Rt ab11143Oxytocin Receptor P IHC-P Rb Hu ab13051

ProteinsOxytocin protein ab9600

Neuroscience > Neuroendocrinology > Thyroid axis


Parathyroid Hormone P IF, IHC-F, IHC-Fr, IHC-P, IP, WB Rb Hu ab7843Parathyroid Hormone P RIA Rb Hu ab14093Parathyroid Hormone P ELISA, RIA Gt Hu ab14163Parathyroid Hormone P ELISA Rb Rt ab14169Parathyroid Hormone P RIA Gt Rt ab14170Parathyroid Hormone [A1/70] M ELISA, IHC-Fr, IHC-P, RIA Mm Hu ab14493Parathyroid Hormone Receptor 1 P IHC-P Rb Hu ab13078Parathyroid Hormone Receptor 1 [3D11] M IHC-P, WB Mm Hu ab3271Parathyroid Hormone Receptor 2 P IHC-P Rb Hu ab13080Thyroid Hormone Receptor [2103] M IHC-Fr, IHC-P, IP, WB Mm Hu ab1131Thyroid Hormone Receptor alpha 1 P GSA, ICC, WB Rb Ch, Hu ab5621Thyroid Hormone Receptor alpha 1+2 P IHC-P Rb Hu, Rt, Pi ab15543Thyroid Hormone Receptor alpha 1+2 [1718] M IHC-P, WB Mm Hu, Mm, Rt, Pi ab16846Thyroid Hormone Receptor beta P GSA, ICC, WB Rb Hu, Rt ab5622Thyroid Hormone Receptor beta P IHC-P Rb Hu, D ab15545Thyroid Hormone Receptor beta [SPM222] M IHC-P Mm Hu, D ab17898Thyroid Hormone Receptor beta 1 [J52] M GSA, ICC, IP, WB Mm Hu, Rt ab2744Thyroid Peroxidase [6H7] M AP Mm Hu ab10246Thyroid Peroxidase [MoAb47] M IHC-P Mm Hu ab12500TSH P ELISA Gt Hu ab8576TSH P IHC-Fr, IHC-P Rb Hu ab9382TSH P RIA Gt Hu ab20085TSH [QB2/6], prediluted M IHC-Fr, IHC-P Mm Hu ab918TSH [TSH-51] M ELISA, ICC, RIA Mm Hu ab7906TSH alpha [ME132 ] M ELISA Mm Hu ab9510TSH beta P ELISA Gt Hu ab21023TSH beta P ELISA Gt Hu ab20625TSH beta [ME-130] M ELISA Mm Hu ab9678TSH beta [ME-130] M ELISA, IA Mm Hu ab9390TSH Receptor P IHC-P Rb Hu ab13136TSH Receptor P IHC-P Rb Hu ab13137TSH Receptor [28] M ELISA, WB Mm Hu, Pi ab2812TSH Receptor [A10] M ELISA, IHC-Fr, IP, WB Mm Hu ab6048

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Neuroscience > Neuronal Markers > Glia > Astrocyte


Astrocyte [1B637] M IHC-FrFl Mm Hu, Rt ab14382Astrocyte [ASTRO1 (10E4/R5)] M IHC-P Mm Hu ab3268GFAP P IF, IHC-F, IHC-P, IP, PCC, WB Rb Ma ab7260GFAP [2A5] M IF, IHC-F, IHC-P, PCC, WB Mm Ma ab8136GFAP [GF-01] M ICC, IHC-Fr, IHC-P, IP, WB Mm Hu, F, Pi.

Does not react with Mm or Rt ab7806GFAP (phospho T7) [YC10] M ICC, WB Mm Hu, C, Pi.

Does not react with Mm or Rt ab12340

S100 antibody (ab7853)

IHC - ab7853 (1/1000) staining of S100 on 30µm coronal rat brain sections. Image shows neuronal cell body andprocesses.

Clonality Applications tested Host Species reactivity DatasheetP IF, IHC-P, IHC-F, IHC-Fr Rb Hu, Rt and Cow www.abcam.com/ab7853

GFAP antibody [6F2] (ab8975)

EM - GFAP (1/100) immunogold labeling of TEM corticobasal degeneration in human brain tissue showing heavy andhighly-specific labeling over a glial process.

Clonality Applications tested Host Species reactivity DatasheetM EM, IHC-P, IHC-Fr, WB Mm Hu www.abcam.com/ab8975

GFAP antibody (ab4674)

IF - ab4674 staining rat astrocytes at a dilution of 1:50 (red). Hoechst dye reveals nuclear DNA in blue.

Clonality Applications tested Host Species reactivity DatasheetP IF, IHC-Fr Ch Hu, Mm, Rt, Ca, C www.abcam.com/ab4674

GFAP antibody (ab7779)

IHC - ab7779 (1/1000) staining GFAP in astrocytes on coronal rat brain tissue sections.

Clonality Applications tested Host Species reactivity DatasheetP IF, IHC-Fr, IHC-P, IHC-F Rb Hu, Rt, Cow www.abcam.com/ab7779

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S100 P IF, IHC-P Rb Hu, Rt, Mm, C, Pi ab868S100 [4E3 ] M IA, P, WB Mm Hu ab10202S100 [6G1] (HRP) M ELISA, IA, P, WB Mm Hu ab10203S100 [8B10] M EIA, ELISA, IF, IHC-P, WB Mm Hu, Rt ab8330S100 [B321] M IF, IHC, WB Mm Hu, Rt, Mm, C, Pi ab7852S100 [B321] (FITC) M IF, IHC-Fr, IHC-P Mm Hu, B ab8570S100A8 + S100A9 [27E10] M FACS, IA, IHC-Fr, IHC-P, IP, WB Mm Hu, Rhesus Mk ab17050

Neuroscience > Neuronal Markers > Glia > Microglia


CD11b antibody [44] M AF, FACS, FuncS Mm Hu ab19674CD11b antibody [44] M AF, FACS, FuncS Mm Hu ab19668CD11b antibody [ICRF44] M FACS, IF Mm Hu ab11355CD11b antibody [LT11] M IA Mm Hu ab10324CD11b antibody [M1/70.15] M FACS Rt Mm ab22371CD11b antibody [M1/70.15] (PE) M FACS, IHC-P Rt Mm ab24518CD11b antibody [M1/70.15] M FACS, ICC, IHC-F, IHC-P, P Rt Hu, Mm, Rb ab6332 Pep. avail.CD11b antibody [M1/70] M BL, FACS, IHC-Fr Rt Mm ab19628CD11b antibody [MEM-174] M FACS, FuncS, IP, WB Mm Hu ab1046CD11b antibody [OX42] M FACS, IHC-Fr, IHC-P, IP Mm Rt ab19576CD11b/c equivalent antibody [OX42] M FACS Mm Rt ab21568CD11b/c equivalent antibody [OX42] M ELISA, FACS, IHC-P Mm Rt ab1211CD11b/c equivalent antibody [OX42] M FACS, IHC-P Mm Rt ab1212CD45 antibody P WB Rb Hu ab10558CD45 antibody [13/2.3] (Biotin) M FACS, IHC-Fr Rt Mm ab19592CD45 antibody [GA90] (FITC) M FACS Mm Hu ab1175CD45 antibody [IBL-3/16] M FACS, IHC-Fr, IP Rt Mm.

Does not react with Rt or Hu ab23910CD45 antibody [K252.1E4] M FACS, IHC-Fr Mm Pi ab23918CD45 antibody [L12/201] M FACS, IHC-Fr, IP Mm Rb ab19566CD45 antibody [MEM-28] M FACS, IF, IHC, IHC-P, IP, WB Mm Hu ab8216CD45 antibody [MRC OX-1] (PE) M FACS Mm Rt ab22269CD45 antibody [MRC OX-1] (FITC) M FACS Mm Rt ab22349CD45 antibody [PD7/26/16 + 2B11] M IHC, IHC-Fr, IHC-P, IM Mm Hu ab781CD45 antibody [YKIX 716.13] M FACS, IP Rt D ab23641CD45 antibody [YW 62.3] (FITC) M FACS Rt Mm ab22475CD45 antibody [YW 62.3] M FACS, IHC-Fr, IP Rt Rt ab22479CD45 peptide M BL ab17550CD45 peptide M BL ab17553HLA DR + DP + DQ antibody [CR3/43] M IHC-P Mm Hu ab17101HLA DR + DP + DQ antibody [WR18] (PE) M FACS Mm Hu ab23901MRP8 antibody [MRP8 7C12/4] M ELISA, FACS, WB Mm Hu ab20220pan Macrophage antibody M IHC-Fr, IHC-P Mm Rt ab15637pan Macrophage antibody [BM8] M FACS, IHC-Fr, IHC-P Rt Hu, Mm ab15694pan Macrophage antibody [RM-1] M FACS, IHC-Fr Mm Rt ab15615

Neuroscience > Neuronal Markers > Glia > Oligodendrocyte


CNPase [11-5B] M Dot, ELISA, ICC, IHC-Fr, Mm B, D, Hu, Mm, Pi, Rb, Rt, Sh. ab6319 Pep. avail.IHC-P, IP, WB Does not react with Ch or Gp

MOSP [CE1] M IF, IHC-Fr Mm Hu, Rt, Mm, F, Ch, Mk, Ca ab3094 Pep. avail.Myelin [2B5] M ICC, IHC-F, IHC-Fr, IHC-P Mm Hu ab8053

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Myelin Basic Protein P WB Rb Hu, Mm, Rt ab12355Myelin Basic Protein P IF, IHC-Fr, IHC-P Rb Hu, Rt ab2404Myelin Basic Protein [12] M AF, ELISA, IF, IHC-Fr, PCC, RIA, WB Rt Hu, B, Gp, Mm, Rb, Rt, Sh ab7349Myelin Basic Protein [B505] M IHC-Fr, IHC-P Mm Hu, Rb, Rt, C, Mk ab8764Myelin Basic Protein [Clone 1 (129-138)] M ELISA, IHC-Fr Mm Hu, Rt, B.

Does not react with Gp ab11160Myelin Basic Protein [Clone 2] M ELISA, IHC-Fr Mm Hu, Rt, B, Gp, Rb, Sh ab11159Myelin PLP [plpc 1] M IF, IHC-Fr, WB Mm Ma ab9311NRG1 P WB Rb Hu, Mm ab2994NRG1 [7D5] M IHC-Fr, IHC-P, IP, WB Mm Rt, Mm ab2369Oct4 P IHC-Fr, IP, WB Rb Hu, Mm, Rt ab18976 Pep. avail.Oligodendrocyte Myelin Glycoprotein [F3-87-8] M IHC-Fr, IP Mm Hu, Rt, D ab24022Oligodendrocyte Specific Protein OSP P IHC-P, WB Rb Mm, Rt ab7474PMP22 P IHC-P Rb Hu ab15506PMP22 [20-14] M IHC-P Mm Hu ab3278Survivin P WB Rb Hu ab8228Survivin [321] M IF, IP, WB Mm Hu ab10584Survivin [6011] M IF, WB Mm Hu, Mm, Rt ab4470Survivin [8E2] M IF, IHC-P Mm Hu, Rt ab3258Survivin (phospho T34) P WB Rb Hu ab10720Survivin peptide BL ab7866Survivin peptide BL ab7865

ProteinsMyelin Basic Protein protein (Biotin) ab792

Neuroscience > Neuronal Markers > Neural Stem Cell


Aggrecan ARGxx [BC-3] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, B, F, D, Gp, H, Pi, Rb, Rt, Sh ab3773BMP2 P Dot, ELISA, WB Rb Hu, Mm, Rt ab14933BMP2 P ELISA, WB Rb Hu ab17597BMP2 [65529.111] M I-ELISA, WB Mm Hu ab6285CD24 [79] M FACS, IHC Rt Mm ab19582CD24 [ALB9] M ICC Mm Hu ab19704CD24 [SN3] M IHC-Fr Mm Hu ab3274CD132 P ELISA, ICC, IF, WB Rb Hu ab16518CD133 M ELISA, WB Mm Hu ab5558CNTF P ELISA, WB Rb Rt ab12426CNTF P I-ELISA, Neut, WB Gt Rt ab10834 Pep. avail.CNTF P ELISA, Neut, WB Rb Rt ab9785 Pep. avail.CNTF P ELISA, IHC, WB Ch Hu, Mm, Rt ab17692CNTF [AS23] M ELISA, IP, WB Mm Hu ab17284

Survivin antibody (ab469)

ICC - HeLa cells in prometaphase, metaphase and anaphase stained with anti-Survivin (green), anti-tubulin (red) andDAPI (blue).

Clonality Applications tested Host Species reactivity DatasheetP WB, IP, IHC-F, IHC-P, H, AF, Rb Ca, D, Hu, Mm www.abcam.com/ab469

IHC-Fr, IF, ICC, IHC

Neuronal m

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Neuroscience > Neuronal Markers > Glia > Oligodendrocyte


34

EMX2 P IHC, WB Rb Hu, Mm, Rt ab11849Integrin beta 1 [4B7] M FACS, H, IA, IF, IHC, IHC-Fr,

IHC-P, IP, WB Mm Hu ab35 Pep. avail.Integrin beta 1 [4B7R] M FACS, IF, IHC-P Mm Hu, Pi ab3167 Pep. avail.Integrin beta 1 [HM beta 11] (Biotin) M FACS Ha Mm ab23825Integrin beta 1 [HM beta 11] M FACS, IP Ha Mm ab23833Integrin beta 1 [HM beta 11] (FITC) M FACS Ha Mm ab23834Integrin beta 1 [MEM-101A] (FITC) M FACS Mm Hu, Mm, Pi ab21845Integrin beta 1 [MEM-101A] M FACS, IP, WB Mm Hu.

Does not react with C ab8238Integrin beta 1 (phospho T788 + T789) P WB Rb Hu ab5189Nestin [10C2] M IF, IHC-Fr, IHC-P, WB Mm Hu. ab22035Nestin [2C13A11] M FACS, ICC, IP, WB Mm Hu.

Does not react with Rt ab18102Nestin [2Q178] M EM, ICC, IHC-Fr, IHC-P, WB Mm Mm, Rt.


IHC (aldehyde-based fixation), IP, WB Mm Hu ab6320PAX2 P IHC-P, WB Rb Hu ab23799SOX2 P IF Rb Hu, Mm ab15830SOX22 P IHC, IP, WB Rb Hu, Mm, Rt ab12062Vimentin P IF Ch Hu, Mm, Rt ab16521

Vimentin antibody (ab7783)

IHC-P - labeling in immunoreactive dermal cells in skin of Balb/c mice using ab7783 in 1% bovine serum albumin for30 min at 37°C.

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-P, IHC-Fr Rb Hu, Mm, Rt www.abcam.com/ab7783

Nestin antibody (ab5968)

ICC - ab5968 (1/200) detecting Nestin immunoreactivity in human embryonic stem cells differentiated in conditionspromoting neuronal differentiation.

Clonality Applications tested Host Species reactivity DatasheetP ICC, IF Rb Hu, Mk. Does not react with Mm www.abcam.com/ab5968

Integrin beta 1 (phospho S785) antibody (ab5185)

ICC - ab5185 staining showing integrin 1 beta (1/5000) in E11 cortical cells cultured for 6 days. Co-staining for beta IIItubulin (not shown) occurs.

Clonality Applications tested Host Species reactivity DatasheetP WB, IF, ICC, IHC Rb Hu, Ch, Mm www.abcam.com/ab5185

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Vimentin P ICC, IF, WB Gt Hu ab11256Vimentin P IF Ch Rt ab24525Vimentin [SP20] M IHC-P Rb Hu ab16700Vimentin [V9] M FACS, IHC-F, IHC-Fr, IHC-P, WB Mm Hu, Rt, F, Ch, D, H, Pi.

Does not react with Mm ab8069Vimentin [VI-01] M ELISA, ICC, WB Mm Ma ab7752Vimentin [VI-10] M ICC, IP, WB Mm Hu, Mm, Rt, Ch, Pi ab20346Vimentin (phospho S55) [4A4] M ELISA, ICC, IHC, WB Mm Hu, Mm, Rt ab22651von Willebrand Factor P IF, IHC-P, IHC-Fr, IM Rb Gp, Hu, Mm, Rt.

Does not react with Ch ab6994

ProteinsCNTF protein ab9786

Neuroscience > Neuronal Markers > Neuron > Dendrite


Drebrin P WB Rb Rt, Hu ab11068Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350MAP2 [AP-20] M ICC, WB Mm Mm, Rt, X, B, Q, S ab11268MAP2 [HM-2] M ICC, IHC, WB Mm Hu, Mm, Rt, B, Ch, Q ab11267

MAP2 antibody [MT-01] (ab7756)

IHC - ab7756 at a dilution of 1/1000, staining MAP-2 on rat brain tissue (30µm thick coronal sections). Labeling ofMAP-2 on cortical neuronal processes/axons.

Clonality Applications tested Host Species reactivity DatasheetM WB, IP, IF, ICC Mm Mm, Ca, C, Pi www.abcam.com/ab7756

MAP2 antibody (ab5392)

IF - ab5392 at a dilution of 1/10000, staining MAP 2 (green) in tissue cultured rat cortical neurons. Nuclei are stainedblue with DAPI.

Clonality Applications tested Host Species reactivity DatasheetP WB, IF, IHC-FrFl Ch C, Hu, Mm, Rt www.abcam.com/ab5392

Vimentin antibody [RV202] (ab8978)

IHC-Fr - vimentin staining of a tonsilar lymphoma with ab8978. The epithelium (on the left) is negative.

Clonality Applications tested Host Species reactivity DatasheetM WB, IHC-Fr, ICC, FACS, IF Mm Hu, Ch, D, Gt, Ha, Mk, www.abcam.com/ab8978

Mm, Rt, C

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MAP2a + MAP2b [AP20] M ICC, IF, IHC-P, WB Mm Hu, Rt, Mm, B, Ch, X , Q ab3096MAP2a + MAP2b [MT-01] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, F ab8479MAP5 [3G5] M IF, IHC-P, IHC on primary cultures Mm Hu, Mm, Rt, C.

Does not react with Ch ab3095neuron specific beta III Tubulin [TU-20] M ICC, IF, IHC-P, IP, WB Mm Ma ab7751neuron specific beta III Tubulin [TUJ-1] M ICC, IF, IHC-Fr, WB Mm Hu, Rt ab14545SAP102 P WB Gt Mm ab12086SAP102 P IHC-Fr, WB Rb Rt.

Does not react with Mm ab3438

Neuroscience > Neuronal Markers > Neuron > Growth Cone


Agrin [AGR 131] M ICC, IHC, IRMA, WB Mm Mm, Rt ab12362Agrin [Agr 247] M ICC, IF Mm Mm, Rt ab12364Agrin [Agr 86] M FACS, ICC, WB Mm Mm, Rt ab12361Axonal Growth Cones [SPM163], prediluted M IHC-P Mm Hu ab17804BAI1 associated protein 2 isoform 3 P WB Gt Hu ab791BAIAP2 P WB Gt Hu ab15697CRMP5 [CR-1] M IHC-Fr, WB Rt Hu, Rt, B ab19378CRMP5 [CR-3] M IHC-Fr, WB Rt Hu, Mm, Rt, B ab19352Doublecortin P IHC-Fr Gp Hu, Mm, Rt ab10293Drebrin P WB Rb Rt, Hu ab11068Drebrin [M2F6] M IHC-Fr, IHC-P, WB Mm Hu, Mm, Rt, Ch, F, Gp, Mk ab12350EphA1 P I-ELISA, WB Gt Hu ab7036EphA1 P ELISA, WB Rb Hu ab5385EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387EphA3 P I-ELISA, WB Gt Mm ab7038EphA3 [III-A4] M FACS, IP Mm Hu ab19439EphA4 P I-ELISA, WB Gt Mm ab7039EphA4 P ELISA, WB Rb Hu ab5389EphA5 P ELISA, WB Rb Hu, Mm ab5397EphA5 P ELISA, WB Gt Mm, Rt ab10612EphA6 P ELISA, WB Gt Mm ab10613EphA7 P ELISA, WB Rb Hu ab5411EphA7 P ELISA, WB Gt Mm ab10614EphA8 P ELISA, WB Gt Mm ab10615EphB1 P I-ELISA, WB Gt Rt ab7042EphB1 P ELISA, WB Rb Hu, Mm ab5414EphB1 P Dot, IP Sh Hu ab10406EphB2 P ELISA, WB Rb Hu, Mm ab5418EphB2 P Dot, ELISA Gt Mm ab10616EphB3 P ELISA, WB Gt Mm ab10617EphB4 P Dot, ELISA, Inhib Gt Mm ab10618EphB6 P Dot, ELISA Gt Mm ab10619Ephrin A2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610Ephrin A3 P ELISA, WB Gt Hu.

Does not react with Mm ab10611Ephrin A4 P I-ELISA, WB Gt Hu ab7041Ephrin B3 P ELISA, WB Gt Hu ab7044GAP43 P H, ICC, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch.

Does not react with X ab7462GAP43 P ICC, IHC-Fr, IP, WB Rb Hu, Mm, Rt, F, Fi, Mk ab12274GAP43 (phospho S41) P WB Rb Rt ab3909Growth Cone [2G13] M IHC-Fr, IHC-P, WB Mm Rt, Mm, Ch ab7762NCAM [123C3] M IF, IHC-F, IHC-Fr, IHC-P Mm Rt, Hu ab8077

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NCAM [B332 (123A8)] M FACS, IHC-Fr, WB Mm Hu ab8596NCAM [ERIC-1] M ELISA, IHC-Fr, IP, RIA, WB Mm Hu ab6123NCAM [H28-123] M AP, IHC-Fr, IM, WB Rt Mm ab19782NCAM [MEM-188] M FACS, IP, WB Mm Hu ab8233NCAM [RNL-1] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt ab9018Neuroserpin P IHC, WB Rb Hu ab16171NRP2 P WB Ch Hu ab15874p35 P WB Rb Hu ab10570p35 [ES19] M IP Mm Hu ab3222p35 nck5a [SPM430] M IP Mm Hu ab19933Robo4 P WB Rb Hu, Rb, Sh ab10547

ProteinsRobo4 protein ab19112

Neuroscience > Neuronal Markers > Neuron > Soma


70 kDa Neurofilament Light P IHC-F, IHC-Fr, IHC-P, WB Rb Ma, Av ab903570 kDa Neurofilament Light [2F11] M IHC-Fr, IHC-P, WB Mm Hu, Op ab897070 kDa Neurofilament Light [DA2] M ICC, IF, IHC-Fr, WB Mm Hu, Mm, Rt, Ch, C, Pi ab4572160 kDa Neurofilament Medium P IF, IHC-F, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, F, B, Pi, Re, Av ab9034160 kDa Neurofilament Medium [3H11] M IHC-F, IHC-Fr, IHC-P, WB Mm Hu ab7256160 kDa Neurofilament Medium [NF-09] M ELISA, ICC, IF, IHC-Fr, IHC-P, WB Mm Ma ab7794160 kDa Neurofilament Medium [RNF403] M IHC-Fr, WB Mm Hu, Rt ab9271200 kDa Neurofilament Heavy P IF, IHC-Fr, IHC-P, WB Rb F, B, Hu, Mm, Pi, Rt ab8135200 kDa Neurofilament Heavy [NAP4] M ELISA, IHC-F, IHC-Fr, IHC-P, WB Mm Ma ab7257200 kDa Neurofilament Heavy [RNF402] M IHC-Fr, IHC-P, WB Mm Hu, Rb, Rt, Mm, B, D, Sh, Gp,

Ch, Ha, Mk, X ab3966200 kDa Neurofilament Heavy [SPM203] M IHC-P, WB Mm Hu ab17879200 kDa + 70kDa Neurofilament [2F11] M IF, IHC, IHC-P, IP, WB Mm Hu ab17114ALK c [ALKc] M IHC-Fr, IHC-P Mm Hu ab650

BrdU [BU1/75 (ICR1)] (ab6326)

Astrocytes from adult (in the lesioned cerebellum) were stained with ab6326. (Green: BrdU; Red: Vimentin).

Clonality Applications tested Host Species reactivity DatasheetM FACS, IHC-P, IHC-Fr Mm Many www.abcam.com/ab6326

BrdU (ab1893)

IF - Tissue culture cells were labelled prior to transplantation and then identified in in vivo tissue using ab1893(10 ug/mL) with a TRITC conjugated secondary antibody.

Clonality Applications tested Host Species reactivity DatasheetP ELISA, IHC-P, IHC-Fr, IP Sh Many www.abcam.com/ab1893

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Neuroscience > Neuronal Markers > Neuron > Growth Cone


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Ataxin 7 P WB Rb Hu ab11434Choline Acetyltransferase P IHC-F, WB Sh Hu, Rt, Rb, Gp, Pi ab18736Choline Acetyltransferase P ELISA, IHC Rb Mm, Pi, Rt ab6168CNS gp130 [F3-87-8] M IHC-Fr Mm Mm ab9310 Pep. avail.Coilin [Pdelta] M ICC, WB Mm Hu.

Does not react with Mm ab11822Doublecortin P IHC-Fr Gp Hu, Mm, Rt ab10293ELAVL2 + ELAVL3 + ELAVL4 [16A11] M IHC-Fr, WB Mm Hu, Mm, Rt, Fi ab14370ELAVL4 [16C12] M IHC-Fr, IHC-P, WB Mm Hu ab14369GAP43 P H, ICC, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch.

Does not react with X ab7462GAP43 P ICC, IHC-Fr, IP, WB Rb Hu, Mm, Rt, F, Fi, Mk ab12274GAP43 (phospho S41) P WB Rb Rt ab3909GARP (truncated) P WB Rb Rt, B ab15094HB9/HLXB9 P EMSA, IHC, IP, WB Rb Hu ab12028MAP2 P IF, IHC, WB Ch B, Hu, Mm, Rt ab5392MAP2 [AP20] M WB Mm Hu, Mm, Rt, C, Ch ab12342MAP2 [HM-2] M ICC, IHC, WB Mm Hu, Mm, Rt, B, Ch, Q ab11267MAP2 [MT-01] M ICC, IF, IP, WB Mm Mm, F, C, Pi ab7756MAP2a + MAP2b [AP20] M ICC, IF, IHC-P, WB Mm Hu, Rt, Mm, B, Ch, X , Q ab3096 Pep. avail.MAP2a + MAP2b [MT-01] M ELISA, IHC-Fr, IHC-P, WB Mm Hu, Mm, F ab8479Necdin P WB Rb Hu, Mm, Rt ab18554Nestin P ICC, IF Rb Does not react with Mm ab5968Nestin [10C2] M IF, IHC-Fr, IHC-P, WB Mm Hu ab22035Nestin [2C13A11] M FACS, ICC, IP, WB Mm Hu.

Does not react with Rt ab18102Nestin [2Q178] M EM, ICC, IHC-Fr, IHC-P, WB Mm Mm, Rt.


IHC (aldehyde-based fixation), IP, WB Mm Hu ab6320neuron specific beta III Tubulin [TU-20] M ICC, IF, IHC-P, IP, WB Mm Ma, Fi ab7751neuron specific beta III Tubulin [TUJ-1] M ICC, IF, IHC-Fr, WB Mm Hu, Rt ab14545

MAP5 antibody [3G5] (ab3095)

IHC-P - High definition dendritic staining with ab3095 (very limited cell soma staining is seen).

Clonality Applications tested Host Species reactivity DatasheetM IHC-fr, IF Mm Hu, Mm, Rt, C. www.abcam.com/ab3095

Does not react with Ch.

c-fos antibody (ab5794)

ICC - ab5794 detecting nuclear staining of c-Fos (green) in adult dorsal root ganglion cells cultured for 2 days.ab5794 was used at 1/100 (incubated for 2 hours at room temperature).

Clonality Applications tested Host Species reactivity DatasheetP ICC Rb Mm www.abcam.com/ab5794

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NSE P WB Ch Hu, Mm, Rt ab14312NSE P WB Rb Hu, Mm, Rt ab16873NSE P IHC-Fr, IHC-P Rb Hu, B ab834NSE [18H2] M ELISA, IP, WB Mm Hu, Mm.

Does not react with Rt ab21253NSE [37E4] M ELISA, WB Mm Hu, Mm, Rt ab16807NSE [85F11] M ELISA, WB Mm Hu, Mm.

Does not react with Rt ab16808NSE [MIG-N3] M IHC-P, WB Mm Hu ab8197NSE [V1-H14] M IHC-Fr, IHC-P Mm Hu ab790PGP95 P IHC-Fr Gp Hu, Mm, Rt ab10410PGP95 P IF, IHC-Fr, WB Rb Hu, Mm, Rt, Pi ab10404PGP95 P WB Gt Hu ab11145PGP95 P IHC-P Rb Hu ab15503PGP95 [13C4 / I3C4] M IF, IHC-F, IHC-Fr, IHC-P Mm Hu, Rt, Gp ab8189PGP95 [31A3] M ELISA, IHC-P, WB Mm Hu, Rt ab20559Plexin A1 P ELISA, WB Rb Hu, Mm, Rt ab23391SOX2 P IF Rb Hu, Mm ab15830SOX22 P IHC, IP, WB Rb Hu, Mm, Rt ab12062STEP / PTPN5 P WB Rb Hu, Mm ab12151STEP / PTPN5 [23E5] M WB Mm Rt ab16967STEP / PTPN5 peptide (phosphatase domain) BL ab22661Survivin P WB Rb Hu ab8228Survivin P AF, H, ICC, IF, IHC, IHC-F,

IHC-Fr, IHC-P, IP, WB Rb F, D, Hu, Mm ab469Survivin [321] M IF, IP, WB Mm Hu ab10584Survivin [6011] M IF, WB Mm Hu, Mm, Rt ab4470Survivin [8E2] M IF, IHC-P Mm Hu, Rt ab3258Survivin (phospho T34) P WB Rb Hu ab10720Survivin peptide BL ab7865Survivin peptide BL ab7866Tyrosine Hydroxylase P ELISA, IHC-Fr Rb Rt ab6211Tyrosine Hydroxylase P ICC, IP, WB Sh Ma ab113Tyrosine Hydroxylase P ICC, IF, IHC, IHC-Fr, IP, WB Rb Ma ab112Tyrosine Hydroxylase [185] M IHC-Fr, WB Mm Hu, Mm, Rt ab10372 Pep. avail.Tyrosine Hydroxylase [300-108] M ICC, IHC, IP, WB Mm Ma ab111Tyrosine Hydroxylase [GD7] M IF, IHC-Fr, WB Mm Hu, Mm, Rt ab23763Tyrosine Hydroxylase (phospho S19) P IF, IHC-Fr, WB Rb Hu, Mm, Rt ab23784Tyrosine Hydroxylase (phospho S31) P IF, IHC-Fr, WB Rb Mm, Rt, Hu ab23782 Pep. avail.Tyrosine Hydroxylase (phospho S40) P IF, IHC, WB Rb Hu, Mm, Rt, Mk, Pi, Q ab16557

NeuN antibody [4G2] (ab13938)

IHC - ab13938 at a dilution of 1/1000, staining NeuN on rat brain tissue (30µm thick coronal sections). Imagesshowing neuronal staining.

Clonality Applications tested Host Species reactivity DatasheetM WB, IHC-P, IHC-Fr, IF, ICC Mm Mm, Rt www.abcam.com/ab13938

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Neuroscience > Neuronal Markers > Neuron > Synapse


14-3-3 P IP, WB Rb Hu, Mm, Rt ab906314-3-3 P IHC-P, IP, WB Rb Hu, Mm, Rt ab608114-3-3 [2Q248] M WB Mm Rt ab1412114-3-3 [3F7] M IP, WB Mm Hu ab1410814-3-3 beta P IHC-P, IP, WB Rb Hu, Mm, Rt ab1526014-3-3 beta P WB Rb Hu, Mm, Rt ab1673014-3-3 beta [60C10] M ELISA, WB Mm Hu, Mm, Rt ab1685914-3-3 beta, prediluted P IHC-P Rb Hu, Mm, Rt ab1526214-3-3 beta + epsilon + zeta [3C8] M WB Mm Hu, Mm, Rt ab1234614-3-3 beta + zeta P WB Rb Ma, Dm ab1411314-3-3 beta + zeta [22-IID8B] M ICC, IHC, WB Mm Hu, Mm, Rt, B, D, Mk, Pi, Rb, Sh ab1234714-3-3 eta + gamma + zeta P ELISA, WB Rb Many ab1412414-3-3 gamma (acetyl V1) [KC21] M WB Mm Hu ab1700514-3-3 gamma [3F8] M IP, WB Mm Hu ab1411714-3-3 sigma [1N6] M IF, IHC-P, IP, WB Mm Hu ab1412314-3-3 tau [3B9] M IP, WB Mm Hu, Mm, Rt, B ab1043914-3-3 Binding Motif (phospho S) P ELISA, IHC-P, IP, WB Rb Many ab1412714-3-3 Binding Motif (phospho S) [3i2] M ELISA, IP, WB Mm Many ab14126Amphiphysin [8] M IHC, IP, WB Mm Hu, Mm, Rt, B ab12232Amphiphysin [C14-23] M ELISA, ICC, IHC, IP, WB Mm Hu, Mm, Rt ab16770 Pep. avail.C3 P IHC-Fr, WB Rb Mm ab11887C3 P IF Sh Hu ab14396 Pep. avail.C3 P ELISA, WB Ch Hu ab14232C3 P DID Gt Hu ab13329C3 P Ie Gt Hu ab20069 Pep. avail.C3 P IF Gt Gp ab20423C3 P ELISA, ID, WB Rb Hu, Mm, Rt ab23891C3 [004-01] M ELISA, WB Mm Hu ab17455C3 [118-02] M ELISA Mm Rt ab17456C3 [11H9] M FACS, IHC-Fr, IP, WB Rt Mm ab11862Calpastatin P ELISA, WB Ch Hu ab16423Calpastatin [2G11D6] M ICC, WB Mm Hu, Rt, C, Pi ab3515CASK P IHC, WB Rb Hu, Rt, Mm ab3383CASK P IP, WB Rb Mm, Rt, D ab11343CD172a [BL1H7] M FACS Mm Pi ab23772Cellubrevin P IF, WB Rb Rt.

Does not react with Hu or D ab2102Cellubrevin P IF, WB Rb Hu, Mm

Does not react with Rt or D ab2103CPEB3 P Fast track Rb Fast track ab12024CPEB3 P WB Rb Mm ab10883CSP P IHC, IP, WB Rb Mm, Rt, X, C ab13250Dynamin [0T178] M ICC, IP, WB Mm Hu, Mm, Rt, B ab14447Dynamin [0T179] M ELISA, IHC-P, IP, WB Mm Hu, Mm, Rt, B ab14450Dynamin [D5] M ICC, IF, IP, WB Mm Hu, Mm, Rt ab12429Dynamin (phospho S774) P IF, IHC, WB Sh Hu, Mm, Rt ab18090Dynamin (phospho S778) P WB Sh Hu, Rt ab18101Dynamin 1 P WB Rb Rt ab3456Dynamin 1 [4E67] M IF, IP, WB Mm B, Ch, Hu, Mm, Rt ab14448Dynamin 1 [D5] M IHC, IP, WB Mm Hu, Mm, Rt, C, Ch ab13251Dynamin 2 P WB Rb Hu, Rt, Mm ab3457Dynamin 3 P WB Rb Hu, Rt, Mm ab3458Homer (1a+1b+1c) P IHC-Fr, WB Rt Hu, Mm, Rt ab11157Homer (1b+1c) P ELISA, FACS, ICC, IF, IHC-Fr,

IP, RIA, WB Rt Hu, Mm, Rt ab18550LAR P WB Rb Hu ab10557LAR peptide BL ab22637Mint1 P WB Rb Rt ab3448MYRIP P Fast track Gt Fast track ab10149PSD93 P IHC-Fr, WB Rb Rt ab2930

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PSD93 P IHC-P, WB Gt Mm ab12097PSD95 P IHC-P, WB Gt Mm ab12093PSD95 [6G6-1C9] M IF, IP, WB Mm Rt ab2723PSD95 [7E3-1B8] M ICC, IP, WB Mm Mm, Rt, B ab13552PSD95 [OT123] M IHC-Fr, IP, WB Mm Hu, Mm, Rt ab18359Rab4 P ICC, IF, IP, WB Rb Hu, Mm, Rt, Ch, D, Fi, Gp,

Ha, Mk, Pi, Rb, Sh ab13252Rab5 P ICC, IHC, IP, WB Rb Hu, Mm, Rt, B, D, Gp, Ha, Mk ab13253Rabphilin 3A P WB Rb Mm, Rt, B ab12234rSec6 [9H5] M IHC, WB Mm Hu, Mm, Rt, B, Ch, D, Ha, Mk,

Pi, Rb, Sh ab12235rSec8 [14G1] M IP, WB Mm Hu, Mm, Rt, B, Ch, D, Gp, Ha,

Mk, Pi, Rb, Sh ab13254SAP102 P IHC-Fr, WB Rb Rt.

Does not react with Mm ab3438SAP102 P WB Gt Mm ab12086 Pep. avail.SIRP alpha P WB Rb Hu, Rt, Mm ab8120SIRP alpha P IHC-P, WB Rb Hu, Mk.

Does not react with Mm ab2971SIRP [MRC OX-41 ] M FACS, IHC-Fr, IHC-P, WB Mm Rt ab9295SNAP23 P WB Rb Rt, Mm ab3340SNAP23 P IF, WB Rb Hu, Rt ab4114SNAP23a P WB Rb Hu, B, D, Ha ab13804SNAP25 [4H251] M WB Mm Hu, Mm, Rt, X, B, Ha, In, Pi ab18002SNAP25 [SP12] M ELISA, IHC-P, WB Mm Hu, Ha, Rt, Pi ab11102SNAP91 [2D11] M IHC, IP, WB Mm Hu, Mm, Rt, B ab12237SNAP91 [AP180-I] M AP, ICC, IP, WB Mm Hu, Rt, B ab11329Synapsin I P AF, IF, IHC-F, IHC-Fr, IP, WB Rb B, Gp, Hu, Mm, Rt ab8Synapsin I (phospho S603) P Dot, WB Rb Rt ab13879Synapsin I (phospho S9) P WB Rb Hu, Mm, Rt, X , B ab16554Synapsin II M IP, WB Rb Mm, Rt, B ab12240Synapsin II P WB Rb Mm, Rt ab13258Synaptobrevin [SP10] M WB Mm Hu, Rt, Ha, Pi ab11109

SNAP25 antibody (ab5666)

IF - SNAP-25 immuno staining in cultured rat hippocampal cells using ab5666 (1/100) yields a pattern consistent withcytoplasmic and plasma membrane staining.

Clonality Applications tested Host Species reactivity DatasheetP WB, IF Rb Mm, Rt, C www.abcam.com/ab5666

SAP97 antibody (ab3437)

IF - Free-floating staining of rat brain tissue with ab3437 (used at 1/3000 overnight).

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-Fr, IHC Rb Rt www.abcam.com/ab3437

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Synaptophysin [SP15] M ELISA, IHC-P, WB Mm Hu, Rt, Ha, Pi ab11105Synaptophysin [SVP38] M FACS, ICC, WB Mm Rt ab12353Synaptophysin [SY38] M IHC-Fr, IHC-P Mm Rt, Mm, B ab8049Synaptobrevin [SP11] M ELISA, IHC-P, WB Mm Hu, Pi.

Does not react with Rt ab11104Synaptophysin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7837Synaptophysin P IHC-Fr, IP, WB Rb Hu, Mm, Rt ab14692Synaptophysin [4E206] M IHC, IP, WB Mm Hu, Mm, Rt, B ab18008Synaptophysin [SP11] M IHC-P, WB Rb Hu ab16659Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037Synaptotagmin P IF, WB Rb Hu, Rt ab10104Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259SynCAM P WB Rb Rt ab3910Syndecan P IHC-P Rb Hu ab16037Syndecan [BC/B-B4] M IHC-Fr, IHC-P Mm Hu ab714Syntaxin P ELISA, IP, WB Ch Hu, B, Mm, Pi, Rt ab8038 Pep. avail.Syntaxin P WB Rb Mm, Rt, C ab13262Syntaxin [SP6] M IHC-Fr, IP, WB Mm Hu, Mm, Rt, X, B, F, Ha, Pi ab12236Syntaxin [SP8] M ELISA, IHC-Fr, WB Mm Hu, Rt, Pi, Ha ab257Syntaxin [STX01 (HPC-1)] M IHC-P, WB Mm Hu, Rb, Rt, C.

Does not react with Gp ab3265Syntaxin 2 P IP, WB Rb Hu, Mm, Rt, X, B, Dm, Ha,

Mk, Pi, Rb, Sh ab12369Syntaxin 3 P IF, WB Rb Ma ab4113Syntaxin 13 [15G2] M ICC, WB Mm Mm, Rt, D, Ha ab13261 Pep. avail.VAMP1 P IP, WB Rb Hu, Rt, Mm, D ab3346 Pep. avail.VAMP2 P IP, WB Rb Rt, Mm ab3347VAMP2 P ICC Ch Hu, Mm, Rt ab14279VAMP8 P IF, IM, IP, WB Rb Hu, Rt, D ab6186

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View the synapse.Neuroscience Antibody

Locator atwww.abcam.com/

neuroscience

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Neuroscience > Neurotransmission > Calcium Signalling > Calcium Binding Proteins


Calbindin P ELISA, IF, IHC-P, IP, Radio-IHC, WB Rb Hu, Rt ab11426Calbindin [CL-300] M IF, IHC-Fr, IHC-P Mm Hu, Ch, Fi, Gp, Ha, Mk, Mm,

Rb, Rt, Sh ab9481Calretinin P IP, WB Rb Hu, Mm, Rt ab14689 Pep. avail.Calretinin P IHC-Fr, IHC-P Rb Hu, F, Ch, Mm, Rt, Re ab702Calretinin [CRT01] M IHC-Fr, IHC-P Mm Mm ab3934Frequenin P Dot, ELISA, IHC-F, IM, WB Ch Most Ma ab7623NCS1 P WB Rb Rt ab5807 Pep. avail.Parvalbumin P ELISA, IHC-P, IP, WB Rb Hu, Rt ab11427 Pep. avail.S100 P IF, IHC-P Rb Hu, Rt, Mm, C, Pi ab868S100 P IF, IHC-F, IHC-Fr, IHC-P Rb Hu, Rt, B ab7853S100 [4E3 ] M IA, P, WB Mm Hu ab10202S100 [6G1] M ELISA, IA, P, WB Mm Hu ab10203S100 [8B10] M EIA, ELISA, IF, IHC-P, WB Mm Hu, Rt ab8330S100 [B321] M IF, IHC, WB Mm Hu, Rt, Mm, C, Pi ab7852 Pep. avail.S100A8 + S100A9 [27E10] M FACS, IA, IHC-Fr, IHC-P, IP, WB Mm Hu, Mk ab17050

Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > L-type


Calcium channel L type DHPR alpha 2subunit [20A] M IHC-Fr, WB Mm Hu, Rb, Rt, Mm, Gp ab2864DHPR alpha 1 P IP, WB Sh Rb ab14446DHPR alpha 1 [1A] M IHC-Fr, IP, WB Mm Hu, Rb, Rt, Mm, Gp ab2862pan Calcium Channel P ICC, IP, WB Rb Mm, Rt ab6298

Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > N-type


CACNB3 P IF, IP, WB Rb Rt ab16717pan Calcium Channel P ICC, IP, WB Rb Mm, Rt ab6298 Pep. avail.

Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > P / Q / R -type


pan Calcium Channel P ICC, IP, WB Rb Mm, Rt ab6298

NCS1 antibody (ab18060)

IF - a cross section of an E14 rat spinal cord at the cervical enlargement labeled with chicken anti-NCS-1(ab18060; 1/100).

Clonality Applications tested Host Species reactivity DatasheetP Conjugation, ELISA, IHC, WB Ch Hu, Mm, Rt www.abcam.com/ab18060

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Neuroscience > Neurotransmission > Calcium Signaling > Calcium Channels > Ryanodine Receptors


Ryanodine Receptor [34C] M IHC-Fr, IHC-P, IP, WB Mm Hu, Rb, Rt, Mm, B, D, Sh, Fi, Am ab2868 Pep. avail.Ryanodine Receptor [C3-33] M FACS, IHC-P, IP, WB Mm Rb, Rt, D, Gp, Ch, Fi, Am ab2827

Neuroscience > Neurotransmission > Calcium Signaling > Calmodulin / CaMK


CABP P WB Gt Hu ab2936CaMKI P IP, WB Rb Hu, Mm, Rt ab15810CaMKII P WB Rb Hu, Mm, Rt, Rb, B, Ch, Gp, Ha ab19223CaMKII [6G9] - Azide free M IHC, RIA, WB Mm Mm, Rt, B ab22609CaMKII (phospho T305) P WB Rb Rt ab22183CaMKII alpha P WB Rb Mm, Rt ab3908CaMKII alpha (phospho T286) P WB Rb Mm ab5683CaMKII alpha (phospho T286) P WB Rb Hu ab18317CaMKII alpha (phospho T286) [22B1] M ELISA, IF, IP, WB Mm Rt ab2724CAMKIV P IHC-Fr, WB Rb Hu, Rt, Mm, D, Mk ab3557CAMKIV P IP, WB Rb Rt ab22658CaMKK P WB Rb Hu, Mm, Rt ab22655Neurogranin P ELISA, ICC, IHC, IP, WB Rt Rt, Mm ab23570

Neuroscience > Neurotransmission > Calcium Signaling > IP3R


IP3R1 P IHC, IP, WB Rb Rt, D ab5908

Neuroscience > Neurotransmission > Intracellular Signaling > Adapters


AP3B2 P ICC, IHC Ch Hu, Mm, Rt ab14227CAPON P Fast track Gt Fast track ab3829GABARAP P WB Rb Hu ab1133PATJ P Fast track Gt Fast track ab8225PDZK3 P Fast track Gt Fast track ab6037PSD P Fast track Gt Fast track ab5962PSD93 P IHC-Fr, WB Rb Rt ab2930PSD93 P IHC-P, WB Gt Mm ab12097PSD95 P IHC-P, WB Gt Mm ab12093PSD95 [6G6-1C9] M IF, IP, WB Mm Rt ab2723

CaMKII alpha antibody [6G9] (ab2725)

IF - Cytoplasmic staining with ab2725 in rat dorsal horn of the spinal cord (lamina II).

Clonality Applications tested Host Species reactivity DatasheetM IF, IP, IHC, WB Mm Rt www.abcam.com/ab2725

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PSD95 [7E3-1B8] M ICC, IP, WB Mm Mm, Rt, B ab13552PSD95 [OT123] M IHC-Fr, IP, WB Mm Hu, Mm, Rt ab18359SAP102 P WB Gt Mm ab12086SAP102 P IHC-Fr, WB Rb Rt.

Does not react with Mm ab3438SAP97 P IHC-Fr, WB Rb Rt ab3437SHANK1 P Fast track Gt Fast track ab10006SHANK1 P ELISA Rb Rt ab18001SHANK1 P WB Rb Hu ab3896SHANK1a P IF, WB Rb Hu ab3897SNTG1 P Fast track Gt Fast track ab10079SynGAP P ICC, WB Rb Rt, Mm ab3344SynGAP P ICC, IP, WB Rb Rt ab17914

Neuroscience > Neurotransmission > Intracellular Signaling > Cytoskeletal


CR16 P WB Gt Mm ab2238Neurabin 1 P IF, WB Rb Hu, Rt ab3483Neurabin 2 P IP, WB Rb Hu, Mm, Rt, Mk, D ab18561Neurofibromin P IP, WB Rb Hu ab17963Neurofibromin [McNFn27] M AP, ELISA, IHC-P, IP, WB Mm Hu, Mm, Rt ab8130Neurofibromin [PcNFn27] P IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt ab8132SNTG1 P Fast track Gt Fast track ab10079

Neuroscience > Neurotransmission > Intracellular Signaling > Kinases


ALK c [ALKc] M IHC-Fr, IHC-P Mm Hu ab650Casein Kinase 1 delta P ELISA, IHC-P Gt Hu ab10877CASK P IHC, WB Rb Hu, Rt, Mm ab3383CASK P IP, WB Rb Mm, Rt, D ab11343 Pep. avail.COPS3 P WB Rb Hu ab10463COPS4 [BL1061] P WB Rb Hu ab12322 Pep. avail.CrkL P WB Gt Hu, Mm ab10907CSN1 P WB Rb Hu ab10413CSN2 P WB Rb Hu ab10426CSN5 [BL1062] P WB Rb Hu ab12323CSN7b P WB Rb Hu ab11895EphA1 P ELISA, WB Rb Hu ab5385EphA1 P I-ELISA, WB Gt Hu ab7036EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387EphA3 P I-ELISA, WB Gt Mm ab7038EphA3 [III-A4] M FACS, IP Mm Hu ab19439EphA4 P ELISA, WB Rb Hu ab5389EphA4 P I-ELISA, WB Gt Mm ab7039EphA5 P ELISA, WB Rb Hu, Mm ab5397EphA5 P ELISA, WB Gt Mm, Rt ab10612EphA6 P ELISA, WB Gt Mm ab10613EphA7 P ELISA, WB Rb Hu ab5411EphA7 P ELISA, WB Gt Mm ab10614EphA8 P ELISA, WB Gt Mm ab10615EphB1 P ELISA, WB Rb Hu, Mm ab5414EphB1 P I-ELISA, WB Gt Rt ab7042

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EphB1 P Dot, IP Sh Hu ab10406EphB2 P ELISA, WB Rb Hu, Mm ab5418EphB2 P Dot, ELISA Gt Mm ab10616EphB3 P ELISA, WB Gt Mm ab10617EphB4 P Dot, ELISA, Inhib Gt Mm ab10618EphB6 P Dot, ELISA Gt Mm ab10619Ephrin A2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610Ephrin A3 P ELISA, WB Gt Hu.

Does not react with Mm ab10611Ephrin A4 P I-ELISA, WB Gt Hu ab7041Ephrin B3 P ELISA, WB Gt Hu ab7044HUNK P ICC, IF, IHC, IHC-P Rb Hu, Mm ab13117Neurogranin P ELISA, ICC, IHC, IP, WB Rt Rt, Mm ab23570

Neuroscience > Neurotransmission > Intracellular Signaling > Phosphatases


PP2A alpha and beta P WB Gt Hu ab1309PP2A alpha and beta P ICC, WB Sh Hu, Mm, Rt, B ab16450PPP2R5A P ICC, WB Rb Hu, Mm, Rt, B ab18136PPP2R5A P WB Gt Hu ab1084PPP2R5A P WB Sh Hu, Mm, Rt ab16449PPP2R5B P WB Gt Mm ab1366PPP2R5C P ICC, WB Rb Hu, Mm, Rt, B ab18133PPP2R5D P WB Gt Hu ab1361PPP2R5E P WB Gt Mm ab1083

Neuroscience > Neurotransmission > Intracellular Signaling > Regulation


CSN1 P WB Rb Hu ab10413 Pep. avail.CSN2 P WB Rb Hu ab10426CSN5 [BL1062] P WB Rb Hu ab12323CSN7b P WB Rb Hu ab11895p35 P WB Rb Hu ab10570p35 [ES19] M IP Mm Hu ab3222p35 nck5a [SPM430] M IP Mm Hu ab19933Phosphoserine P ELISA, IP, WB Rb Many ab9332Phosphoserine (HRP) P ELISA, IP, WB Rb Many ab9334Phosphoserine [3C171] M ELISA, WB Mm Many ab17465Phosphoserine [4A9] M ELISA, IP, WB Mm Many ab21642Phosphoserine [PSR-45] M I-ELISA, WB Mm Many ab6639

Phosphoserine/threonine/tyrosine antibody [SPM101] (ab15556)

IHC - Human breast carcinoma stained with ab15556 (FFPE-sections).

Clonality Applications tested Host Species reactivity DatasheetM WB, IP, ELISA, IHC-P, IF Mm Many www.abcam.com/ab15556

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Phosphoserine/threonine P ELISA, IP, WB Rb Many ab17464Phosphothreonine-Proline /Phosphoserine-Proline P ELISA, WB Rb Many ab9344Phosphothreonine-Proline /Phosphoserine-Proline (HRP) P ELISA, WB Rb Many ab9345Phosphotyrosine P ELISA, WB Rb Many ab17301Phosphotyrosine P Dot, ELISA, IF, WB Rb Many ab17303Phosphotyrosine P ELISA, IP, WB Rb Many ab9329Phosphotyrosine P ELISA, IP, WB Rb Many ab9319Phosphotyrosine (AP) M ELISA, RIA Mm Many ab2339Phosphotyrosine [1BB49] (Biotin) M IHC, WB Mm Many ab17371Phosphotyrosine [1BB49] (FITC) M IHC, WB Mm Many ab17372Phosphotyrosine [2Q258] (HRP) M ELISA, WB Mm Many ab17288Phosphotyrosine [P9V6] (Biotin) M IA, WB Mm Many ab7227Phosphotyrosine [PY20] (Biotin) M ELISA, ICC, WB Mm Many ab16388Phosphotyrosine [IG2] M FACS, ICC, IP, WB Mm Many ab5592Phosphotyrosine [P9V6] M IA, WB Mm Many ab7226

Neuroscience > Neurotransmission > Neurotransmitters > Acetylcholine


Acetylcholine P ELISA, ICC, IHC-P Rb Many ab8884Acetylcholine M ICC, IHC-P Mm Many ab6470Acetylcholinesterase P ELISA, IP, Neut, PCC, WB Gt B ab7396Acetylcholinesterase (HRP) P ELISA, IHC-Fr, IM, PCC, WB Gt B ab7397Acetylcholinesterase [190-01] M ELISA, WB Mm Hu, B ab17774Acetylcholinesterase [HR2] M ELISA, IHC-Fr, IP Mm Hu, Rb, B, F, Gp ab2803Acetylcholinesterase [HYB 111-05] M ELISA, WB Mm Hu.

Does not react with Fi ab23455Acetylcholinesterase [ZR3] M IHC-Fr, IP Mm Rb, Rt, F, Gp ab2802Choline Acetyltransferase P IHC-F, WB Sh Hu, Rt, Rb, Gp, Pi ab18736Choline Acetyltransferase P ELISA, IHC Rb Mm, Pi, Rt ab6168PBP P WB Gt Hu ab2634

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Aspartate


Glutamate Aspartate Transporter P FACS, ICC, IHC, PCC, WB Rb Rt ab416L-Aspartate P ICC, IHC-F, IHC-Fr, IM Rb Many ab9439

Phosphotyrosine antibody [PY20] (ab10321)

IF - in the rat cortex. Shows microglial cells stained. IHC free-floating protocol using 4% PFA fixed brain tissue.Primary antibody ab10321 was used at 1 ug/ml incubated overnight at room temperature.

Clonality Applications tested Host Species reactivity DatasheetM AP, ELISA, FACS, IF, IHC-FrFl, IP, WB Mm Rt www.abcam.com/ab10321

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Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > GABA


GABA A Receptor alpha 1 P IP, WB Rb Rt ab5781GABA A Receptor alpha 2 P WB Rb Mm, Rt ab8342GABA A Receptor alpha 4 P IP, WB Rb Mm, Rt ab4120GABA A Receptor alpha 4 P Dot, WB Rb Rt ab13872GABA A Receptor alpha 5 P WB Rb Rt, Mm ab3700GABA A Receptor alpha 6 P IP, WB Rb Mm, Rt ab5365GABA A Receptor beta 1 P IP, WB Rb Mm, Rt ab16703GABA A Receptor beta 1 P WB Rb Hu ab23338GABA A Receptor beta 2 P IP, WB Rb Mm, Rt ab8340GABA A Receptor beta 3 P IP, WB Rb Mm, Rt ab4046GABA A Receptor delta P WB Rb Hu, Mm ab10100GABA A Receptor gamma 2 P IP, WB Rb Mm, Rt ab4073GABA P ICC, IHC-F, IHC-Fr, IM Rb Many ab9446GABA P IF, IHC Gp Many ab17413GABA P ELISA, IF, IHC Rb Rt ab17390GABA [2Q67] M IHC-Fr, IHC-P Mm Hu ab17313GABA B Receptor 1 P WB Rb Hu ab1135GABA B Receptor 2 (phospho S892) P IF, IHC, WB Rb Mm, Rt ab3900GABA B Receptor 2 P WB Rb Hu ab1134GABA Transporter 1 / GAT 1 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab426GABA Transporter 2 P IF, IHC-Fr, WB Rb Hu, Mm, Rt, Ch ab2896GABA Transporter 3 / GAT 3 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab431

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Glutamate


GAD65 P IHC-Fr, IHC-P Rb Hu, Rt ab2348GAD65, prediluted P IHC-P Rb Hu, Rt ab15299Glutamate P ICC, IHC-F, IHC-Fr, IM Rb Many ab9440Glutamate P ELISA, ICC Rb Many ab8889Glutamate M ELISA, IHC-P Mm Many ab8893Glutamate [2Q71] M IHC Mm Rt ab17463Glutamate Aspartate Transporter P FACS, ICC, IHC, PCC, WB Rb Rt ab416Glutamate Transporter 1 P FACS, ICC, IF, IHC, PCC, WB Rb Hu, Rt ab417Glutamate Receptor 6+7 (metabotropic) P IHC-P, IP, WB Rb Hu, Mm, Rt, Ch, Mk ab15307

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Glycine


Glycine P ICC, IHC-F, IHC-Fr, IM Rb Many ab9442Glycine Receptor alpha 1 P IP, WB Rb Many ab475

Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Related targets


5-hydroxy Tryptamine P ELISA, IHC-Fr Rb Many ab64586-hydroxy Tryptamine P ELISA, IHC-P Rb Many ab6455ARMET P WB Gt Hu ab1347ARMET peptide (222-234) ab22811Taurine P ELISA, IHC-Fr Rb Many ab6467Taurine P ICC, IHC-F, IHC-Fr, IM Rb Many ab9448Tyramine P ELISA Rb Many ab6451

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Neuroscience > Neurotransmission > Neurotransmitters > Amino Acids > Tryptophan


5-hydroxy Tryptophan P ELISA, ICC, IHC-Fr Rb Many ab88905-hydroxy Tryptophan [4D18] M ELISA Mm Many ab181715-methoxy Tryptophan P ELISA Rb Many ab6476Tryptophan P ELISA, ICC Rb Many ab8886Tryptophan Hydroxylase P IF, IHC, WB Sh Hu, Rt ab3907Tryptophan Hydroxylase (phospho S58) P WB Rb Hu, Mm, Rt, X ab23779

Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Dopamine


6-hydroxy Dopamine P ELISA Rb Many ab9429DOPA Decarboxylase P ICC, WB Rb Rt ab3905Dopamine M BL, IHC-Fr, IHC-P Mm Many ab1001Dopamine P ELISA, IHC Rb Many ab6427Dopamine P ELISA, ICC Rb Many ab8888 Pep. avail.Dopamine M ELISA, ICC, IHC-Fr, IHC-P Mm Many ab8892 Pep. avail.Dopamine [2B11] M ICC, Neut Mm Many ab8281 Pep. avail.Dopamine beta Hydroxylase P IHC, WB Sh B, Hu, Rb, Mk.

Does not react with D, Rt or Sh ab6487 Pep. avail.Dopamine beta Hydroxylase P IF, IHC-P, WB Sh Hu, Mm, Rt, B ab19353Dopamine Transporter P ELISA, WB Rb Hu, Mm, Rt ab18548Dopamine Transporter [hDAT-LOOP] M ICC, IHC-Fr, WB Rt Hu, Rt ab5981L Dopa M ELISA Mm Many ab5979L Dopa P ELISA, IHC-Fr Rb Many ab6426Methamphetamine [4E2] M ELISA Mm Many ab23328Neurotensin P IHC-Fr, IHC-P, IHC-R Rb Ma ab11142Tyrosine Hydroxylase (phospho S19) P IF, IHC-Fr, WB Rb Hu, Mm, Rt ab23784Tyrosine Hydroxylase (phospho S31) P IF, IHC-Fr, WB Rb Mm, Rt, Hu ab23782Tyrosine Hydroxylase (phospho S40) P IF, IHC, WB Rb Hu, Mm, Rt, Mk, Pi, Q ab16557Tyrosine Hydroxylase P ICC, IF, IHC, IHC-Fr, IP, WB Rb Many ab112Tyrosine Hydroxylase P ICC, IP, WB Sh Many ab113Tyrosine Hydroxylase P ELISA, IHC-Fr Rb Rt ab6211Tyrosine Hydroxylase [185] M IHC-Fr, WB Mm Hu, Mm, Rt ab10372Tyrosine Hydroxylase [300-108] M ICC, IHC, IP, WB Mm Ma ab111Tyrosine Hydroxylase [GD7] M IF, IHC-Fr, WB Mm Hu, Mm, Rt ab23763

Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Noradrenaline


Noradrenaline P ELISA, IHC-Fr Rb Many ab6454Noradrenaline P ELISA, ICC, IHC-Fr Rb Many ab8887

Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Related targets Amines


5-methoxy Tryptamine P ELISA, IHC-Fr Rb Many ab64586-hydroxy Tryptamine P ELISA, IHC-P Rb Many ab6455CSPS P WB Rb Hu, Mm, Rt ab16870CSPS [3F10] M ELISA, IP Mm Hu, Mm, Rt ab16773Tyramine P ELISA Rb Many ab6451

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Neuroscience > Neurotransmission > Neurotransmitters > Biogenic Amines > Serotoin / 5HT


5 Methoxytryptamine P IHC Rb Rt ab151655HT1B Receptor P WB Rb Hu, Mm, Rt ab13896Nitroserotonin P ELISA Rb Many ab6459Serotonin P IHC-Fr, IHC-P Rb Hu, Rt ab10385Serotonin P ELISA, ICC, IHC-Fr Rb Many ab8882Serotonin [5HT-H209] M ICC, IHC-Fr, IHC-P Mm Hu, Mm, Rt, Gp ab16007Serotonin [YC5/45] M IHC-P Rt Hu, Mm, Rt, Gp, Rb, X ab6336Serotonin N-acetyltransferase P WB Rb Rt ab3505Serotonin N-acetyltransferase (phospho S206) P WB Rb Rt ab3440Serotonin N-acetyltransferase (phospho T29) P WB Rb Rt, Mm ab3439

Neuroscience > Neurotransmission > Neuropeptides > Hormones


ACTH P ICC, WB Ch Hu, Mm, Rt ab14064ACTH P Dot, IHC-Fr Rb C, Rt ab8906ACTH P IHC-P Rb Hu ab10335ACTH P RIA Rb Hu, Rt, Mk ab11116ACTH P IHC-Fr, IHC-P Rb Hu ab2407ACTH [56] M ELISA, ICC, IHC-P Mm Hu, Rt ab21003ACTH [A2H8] M ELISA, RIA Mm Hu ab11114ACTH [A-4D5] M RIA Mm Hu ab11113ACTH [B427] M IF, IHC-F, IHC-Fr, IHC-P, ICC, WB Mm Hu ab8615BNP P ELISA, IHC, RIA Rb Hu, Rt.

Does not react with Mm ab19645BNP P ELISA, IF, RIA Rb Hu ab19646BNP [1B6] M EIA, ELISA, WB Mm Hu ab14699BNP [2d3] (HRP) M EIA, ELISA Mm Hu ab14701CGRP P IHC-F Sh Hu, Rt ab22560CGRP P RIA Gt Hu ab14180CGRP P IHC-Fr, IHC-P Rb Hu, Rt ab8056CGRP [4901] M ELISA, ICC, Neut, RIA Mm Hu, Rt, D ab10987Corticoliberin [4H9] M ELISA Mm Hu ab10034Corticotropin Releasing Factor P AP, ICC, RIA Rb Hu, Rt ab8901Estradiol P RIA Sh Many ab22441Estradiol [6E1] M RIA Mm Hu ab20257Estradiol [B42] M ELISA, IHC-Fr Mm Ma ab8743Exendin 4 [ABS 012-20] M ELISA Mm Re ab23407MC1 Receptor P IHC-P Rb Hu ab13033MCH P ELISA, IHC-Fr Ch Hu, Mm, Rt ab15677MSH gamma 1 P Dot, RIA, WB Rb C, Rt ab8910Neurokinin B P IHC-P Rb Hu, Rt ab5772Neuropeptide EI NH2 P IHC-Fr Rb Rt ab11141Neuropeptide Y P IHC-Fr Gp Rt ab10341Neuropeptide Y P Dot, IHC-Fr, RIA Rb Hu, Mm, Rt, F, B, Mk, Pi, Rb ab10980Neuropeptide Y P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Gp, Rb ab6173Orexin A P ELISA, ICC, IHC-Fr Rb Hu ab16787Orexin A P ELISA, RIA Rb Hu, Mm, Rt ab10981Oxytocin P IHC-Fr Rb Rt ab11143Oxytocin P IHC-P, RIA Rb Hu, Rt, Mm, Rb, Sh ab2078Oxytocin Receptor P IHC-P Rb Hu ab13051Peptide YY P IHC-Fr, IHC-P, IHC-R Rb Pi ab22663Peptide YY P ELISA, WB Ch Hu, Mm, Rt ab15879proBNP P RIA Gt Hu ab14352proBNP [10h4] M EIA, ELISA, WB Mm Hu ab14703proBNP [24E11] (HRP) M ELISA, WB Mm Hu ab13123Somatostatin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7840

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Somatostatin 14 P Dot, WB Rb Rt ab8903Somatostatin 14, prediluted P IHC-P Rb Hu ab15522Somatostatin 28 P EM, IF, IHC-Fr, IHC-P Rb Many ab22682Somatostatin 28 P Dot, RIA, WB Rb Rt ab8904Somatostatin Receptor P IHC-Fr, IHC-P Rb Rt, Mm ab2366Somatostatin Receptor 1 P FACS, WB Rb Hu, Rt ab5827Somatostatin Receptor 1 P IP, WB Rb Hu, Mm, Rt ab479Somatostatin Receptor 2 P IF, IHC-P Rb Hu, Rt ab13120Somatostatin Receptor 2 P IF, WB Rb Hu, Mm, Rt ab9550Substance P P IHC-Fr Gp Hu, Mm, Rt ab10353Substance P P RIA Rb Hu ab7305Substance P P ELISA, WB Rb Many ab1123Substance P [M09205] M IHC-Fr Rt Hu ab7340Substance P [NC1/34 HL] M ELISA, H, ICC, IF, IHC, IHC-F,

IHC-Fr, IHC-P, RIA Rt Av ab6338Substance P [SP-DE4-21] M ELISA, IHC-P Mm Hu, Rt, Gp ab14184Vasopressin P IHC-F, IHC-Fr, IHC-P Rb ab8745VIP P IHC-Fr, IHC-P Rb Hu, Rt, Pi ab22736VIP P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Mm, Rt, B, D, Gp, Pi, Rb ab6184

ProteinsACTH protein ab9586Neuropeptide Y protein ab9599Oxytocin protein ab9600proBNP protein ab13125Somatostatin 28 protein ab9606Somatostatin protein ab9607Substance P protein ab9608VIP protein ab9609

Neuroscience > Neurotransmission > Neuropeptides > More Neuropeptides


FMRFamide P IHC-Fr Rb Rt ab10352Neuropeptide EI NH2 P IHC-Fr Rb Rt ab11141Neuropeptide FF1 P WB Rb Hu ab3898Neuropeptide Y P IHC, IHC-F, IHC-Fr, IHC-P Sh Hu, Gp, Rb ab6173Neuropeptide Y P IHC-Fr Gp Rt P ab10341Neuropeptide Y P Dot, IHC-Fr, RIA Rb Hu, Mm, Rt, F, B, Mk, Pi, Rb ab10980Neuropeptide Y CPON P IHC-P, IHC-R Rb Many ab22695NPFF1 Receptor P ELISA, WB Rb Hu ab1401NPFF1 Receptor P IHC-P Rb Hu ab13367NPFF2 P WB Rb Hu ab3899NPFF2 Receptor P ELISA, WB Rb Hu ab1402NPFF2 Receptor P IHC-P Rb Hu ab13198NPY5R P ICC, IHC-P Rb Hu ab13199Orexin B P ELISA, RIA Rb Hu, Mm, Rt ab10982Orexin B P ELISA, IHC, WB Rb Mm ab14970PBP P WB Gt Hu ab2634VRL1 P Dot, IHC-Fr, WB Rb Rt ab6183

ProteinsNeuropeptide Y protein ab9599

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Neuroscience > Neurotransmission > Neuropeptides > Hormones


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Neuroscience > Neurotransmission > Neuropeptides > Opioids


Endomorphin 1 & 2 P ELISA, IHC-Fr Rb Hu, Mm, Rt, Mk, Pi ab10290Endomorphin 1 P ELISA, IHC-Fr Rb Hu, Mm, Rt, Mk, Pi ab10285Morphine M ELISA Mm Not applicable ab1060Morphine [3A6] M ELISA Mm Not applicable ab23357Nociceptin P IHC-Fr Gp Rt, Mk ab10276Nociceptin P IHC-Fr Rb Rt, Mk ab10277Nociceptin P ELISA, I-ELISA, IHC-Fr, WB Rb Gp, Rt ab6174NPFF2 P WB Rb Hu ab3899Peptide E P IHC-Fr Rb Rt, B ab11144ProDynorphin P IHC-Fr Gp Rt ab10280ProDynorphin P IHC-Fr Rb Mm, Rt ab11137

Neuroscience > Neurotransmission > Neuropeptides > Tachykinins


Neurokinin 1 Receptor NK1 P FACS, WB Rb Hu ab466Neurokinin 1 Receptor NK1 P AF, IHC-F, IHC-Fr Rb Gp, Mm, Rt ab6Neurokinin 1 Receptor NK1 P IHC-Fr, WB Rb Rt, Mm, Gp ab8873Neurokinin 1 Receptor NK1 P IF, IHC-P Rb Hu, Rt.

Does not react with Gp ab13133Neurokinin 3 Receptor NK3 P AF, IHC, IHC-F, IHC-Fr Rb Gp, Mm, Rt ab7Neurokinin 3 Receptor NK3 P IHC-P Rb Hu ab13135Neurokinin A Receptor P ICC, IHC-P Rb Hu ab13397Neurokinin B P IHC-P Rb Hu, Rt ab5772Substance P P ELISA, WB Rb Many ab1123 Pep. avail.Substance P P RIA Rb Hu ab7305Substance P P IHC-Fr Gp Hu, Mm, Rt ab10353Substance P [M09205] M IHC-Fr Rt Hu ab7340Substance P [NC1/34 HL] M ELISA, H, ICC, IF, IHC, IHC-F,

IHC-Fr, IHC-P, RIA Rt Av, Ma ab6338Substance P [SP-DE4-21] M ELISA, IHC-P Mm Hu, Rt, Gp ab14184Thy12 [FF-10] M FACS, IHC-Fr, IP, WB Rt Mm ab22489

ProteinsSubstance P protein ab9608

Neuroscience > Neurotransmission > Nitric Oxide > Nitrated Species


Nitrotyrosine P ELISA, IA, WB Gt Many ab20117Nitrotyrosine P WB Rb Many ab23704Nitrotyrosine P ELISA Ch Many ab2494Nitrotyrosine [HM11] M ELISA, IHC-Fr, IHC-P, WB Mm Hu ab7048NO Tyrosine P ELISA Rt Many ab6479NO Tyrosine P ELISA Rb Many ab3916NO2 Tyrosine P ELISA Rb Many ab6471NO2 Tyrosine P ELISA Rt Many ab3805

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Neuroscience > Neurotransmission > Nitric Oxide > NOS


bNOS P IHC-Fr, WB Rb Rb, Rt, Mm, B ab3511bNOS P IF, IHC-F, IHC-Fr, IHC-P, WB Sh Gp, Rb, Rt ab6175bNOS P WB Rb Hu, Rt ab19892CAPON P Fast track Gt Fast track ab3829DDAH2 P WB Gt Mm ab1383 Pep. avail.eNOS P ELISA, IP, WB Rb B ab3868eNOS P ICC, WB Rb Hu, Mm, Rt, B ab4225eNOS P IHC-P, WB Rb B, D, Hu, Mm, Pi, Rt ab5589eNOS (phospho S116) P ELISA, IP, WB Gt B ab3867iNOS P IHC-P, WB Rb Hu, Mm, Rt ab15323iNOS P IF, WB Rb Hu, Rt, Mm ab3523iNOS P ELISA, WB Rb Hu, Mm, Rt ab4226NOS P IHC-P, WB Rb Hu, Mm, Rt, C, Pi ab15203nNOS (neuronal) P WB Rb Hu ab3895nNOS (neuronal) P WB Rb Rt ab5586nNOS (neuronal) (phospho S1416) P WB Rb Rt ab5583pan NOS P IHC-Fr, WB Rb Hu, B, Dm, Mm, Pi, Rt, Fi ab3342pan NOS [NOS-3F7-B11 B5] M IHC-Fr, WB Mm Rt, Mm, B ab2801

Neuroscience > Neurotransmission > Pharmacological Tools


Amitriptyline [1BB831] M ELISA, RIA Mm Not applicable ab15176Amitriptyline [202] - BSA and Azide free M IA Mm Not applicable ab20408Amphetamine P ELISA, IF, IHC, IP, WB Gt Not applicable ab15208Amphetamine [2DD1] M ELISA Mm Not applicable ab15210Atrazine P ELISA Sh Not applicable ab22435Benzodiazepine P ELISA Sh Not applicable ab15212Ketanserin M ELISA Mm Not applicable ab1061Phenobarbital M ELISA Mm Not applicable ab1097Phenobarbital [F1#2G7A7] M EIA, RIA Mm Not applicable ab20871

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Acetylcholine


Muscarinic Acetylcholine Receptor 1 P WB Rb Mm ab3480Muscarinic Acetylcholine Receptor 2 P IHC-P Rb Hu ab13043Muscarinic Acetylcholine Receptor 2 [31-1D1] M IP, WB Mm Hu, Rt, Mm, Pi.

Does not react with Ch ab2805Muscarinic Acetylcholine Receptor M3 P IHC-P Rb Hu ab13063Muscarinic Acetylcholine Receptor M4 P IHC-P Rb Hu ab13065

nNOS antibody (ab1376)

IHC-FrFl - ab1376 immunostaining nNOS in rat dorsal root ganglion neurons 2 weeks post spinal nerve ligation.Thestaining is restricted to the cytoplasm of a subpopulation of neurons.

Clonality Applications tested Host Species reactivity DatasheetP WB, IHC-FrFl Gt Mm www.abcam.com/ab1376

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Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Adrenergenic Receptors


alpha 1b Adrenergic Receptor P WB Ch Hu ab15851alpha 1b Adrenergic Receptor P IHC-P Rb Hu ab12933alpha 1c Adrenergic Receptor P IHC-P Rb Hu ab12935alpha 1d Adrenergic Receptor P IHC-P Rb Hu ab12923

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Cannabinoid Receptors


Cable pilus P WB Rb Bu ab13961Cannabinoid Receptor 1 P IHC-P, WB Rb Hu, Mm, Rt, C, D ab23703Cannabinoid Receptor I P ICC, WB Rb Hu, Rt ab3558Cannabinoid Receptor I P ICC, IHC-P Rb Hu ab13166Cannabinoid Receptor I P IF, IHC-Fr, WB Rb Hu, Mm, Rt ab10573Cannabinoid Receptor II P ICC, IHC-Fr, WB Rb Hu ab3560Cannabinoid Receptor II P ICC Rb Hu, Rt ab3561

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Dopamine Receptors


DARPP32 P ELISA, IHC-P, WB Rb Hu, Mm, Rt ab18551DARPP32 (phospho T34) P ELISA, WB Rb Hu, Rt ab18553DARPP32 (phospho T75) P ELISA, IHC, WB Rb Hu, Rt ab18552Dopamine Receptor D1 P ICC, IHC-P Rb Hu ab12969Dopamine Receptor D4 P IHC-P Rb Hu ab13318Neurotensin Receptor 1 P IHC-P Rb Hu ab13201Neurotensin Receptor 2 P IHC-P Rb Hu ab13068

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Glutamate


GRM8 P IHC-P Rb Hu ab13194Homer (1a+1b+1c) P IHC-Fr, WB Rt Hu, Mm, Rt ab11158Homer (1b+1c) P ELISA, FACS, ICC, IF, IHC-Fr,

IP, RIA, WB Rt Hu, Mm, Rt ab18550Metabotropic Glutamate Receptor 1 P IHC-P Rb Hu ab13037Metabotropic Glutamate Receptor 1a P IF, IHC, IHC-Fr, IP, WB Rb F, Mm, Rt ab6439Metabotropic Glutamate Receptor 1a P ICC, IHC-Fr, WB Rb Hu, Mm, Rt, F ab10302Metabotropic Glutamate Receptor 2 P WB Rb Rt ab10304Metabotropic Glutamate Receptor 2 P ICC, IHC-P Rb Hu ab13192Metabotropic Glutamate Receptor 2 [mG2Na-s] M IF, IHC, WB Mm Rt, Mm ab15672Metabotropic Glutamate Receptor 2+3 P IHC, IP, WB Rb F, Mm, Rt ab6438Metabotropic Glutamate Receptor 2+3 P ICC, IHC-Fr, WB Rb Rt ab10307Metabotropic Glutamate Receptor 3 P WB Rb Rt ab10309Metabotropic Glutamate Receptor 5 P IHC, IP, WB Rb F, Mm, Rt ab6436Metabotropic Glutamate Receptor 5+1 P IHC-Fr, IP, WB Rb Mm, Rt ab16214Metabotropic Glutamate Receptor 6 P WB Rb Rt ab10314Metabotropic Glutamate Receptor 7 P IHC-P Rb Hu ab13042mGluR5 P WB Rb Mm, Rt ab10311

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Neuroscience > Neurotransmission > Receptors / Channels > GPCR > More GPCR


Adenosine A3 Receptor P IHC-P Rb Hu ab13160APJ Receptor P ICC, IHC-P Rb Hu ab13267APJ Receptor P IHC-Fr Ch Mm, Rt ab15639AVPR1B P IHC-P Rb Hu ab13147AVPR2 P IHC-P Rb Hu ab13149Calcitonin receptor P ELISA, IHC-P, WB Rb Hu, Mm, Rt ab11042CCKBR P IHC-P Rb Hu ab13173CCKBR P WB Rb Hu ab14439Corticotropin Releasing Factor Receptor 2 P IF, IHC-Fr, IHC-P, WB Rb Hu, Rt ab12964CysLT1 P IHC-P Rb Hu ab12968CysLT2 P IHC-P Rb Hu ab13174EDG5 P WB Rb Hu ab12259EDG5 P ICC, IHC-P Rb Hu ab13127EDG7 P ICC, IF, WB Rb Hu, Mm, Rt ab23692EDG7 P IHC-P Rb Hu ab13350GABA B Receptor 1 P WB Rb Hu ab1135GABA B Receptor 2 P WB Rb Hu ab1134GABA B Receptor 2 (phospho S892) P IF, IHC, WB Rb Mm, Rt ab3900GALR2 P IHC-P Rb Hu ab13179GALR3 P IHC-P Rb Hu ab13003Gastric Inhibitory Polypeptide Receptor P IHC-P Rb Hu ab13005GHRHR P IHC-P Rb Hu ab13342GLP1R P IHC-P, WB Rb Hu, Mm, Rt, Ha ab13181HRH3 P ICC, IHC-P Rb Hu ab13014HRH4 P IHC-P Rb Hu ab13183LPHN2 P IHC-P Rb Hu ab13019LPHN3 P IHC-P Rb Hu ab13022MASS1 P IHC-P Rb Hu ab13150MC4 Receptor P ICC, IHC-Fr, IHC-P Rb Hu ab13034 Pep. avail.MC4 Receptor P IHC-P, WB Rb Hu, Mm, Rt ab24233Melatonin Receptor 1A P IHC-P Rb Hu ab13035Melatonin Receptor 1B P IHC-P Rb Hu ab13357Melatonin Related Receptor P IHC-P Rb Hu ab13190Neurokinin A Receptor P ICC, IHC-P Rb Hu ab13397NMUR1 P IHC-P Rb Hu ab13365NMUR2 P IHC-P Rb Hu ab13366NPFF1 Receptor P IHC-P Rb Hu ab13367NPFF1 Receptor P ELISA, WB Rb Hu ab1401NPFF2 Receptor P IHC-P Rb Hu ab13198 Pep. avail.NPFF2 Receptor P ELISA, WB Rb Hu ab1402NPY5R P ICC, IHC-P Rb Hu ab13199OA1 P IHC-P Rb Hu ab13070OR10R2 P IHC-P Rb Hu ab13072OR2H3 P IHC-P Rb Hu ab13073P2Y1 P IHC-P Rb Hu ab13271P2Y10 P IHC-P Rb Hu ab13272P2Y11 P IHC-P Rb Hu ab13104P2Y12 P ICC, IHC-P Rb Hu ab13077 Pep. avail.P2Y2 P IHC-Fr Rb Hu, Mm, Rt ab10270 Pep. avail.P2Y8 P IHC-P Rb Hu ab13273P2Y9 P IHC-P Rb Hu ab13351Parathyroid Hormone Receptor 2 P IHC-P Rb Hu ab13080Somatostatin Receptor 1 P IP, WB Rb Hu, Mm, Rt ab479Somatostatin Receptor 1 P FACS, WB Rb Hu, Rt ab5827Somatostatin Receptor 2 P IF, WB Rb Hu, Mm, Rt ab9550Somatostatin Receptor 2 P IF, IHC-P Rb Hu, Rt ab13120TA4 P IHC-P Rb Hu ab13403

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Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Muscarinic Receptors


Muscarinic Acetylcholine Receptor 1 P WB Rb Mm ab3480Muscarinic Acetylcholine Receptor 2 P IHC-P Rb Hu ab13043Muscarinic Acetylcholine Receptor 2 [31-1D1] M IP, WB Mm Hu, Rt, Mm, Pi.

Does not react with Ch ab2805Muscarinic Acetylcholine Receptor M3 P IHC-P Rb Hu ab13063Muscarinic Acetylcholine Receptor M4 P IHC-P Rb Hu ab13065

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Opioid Receptors


Delta Opioid Receptor P IHC-Fr Rb Mm, Rt, Primate ab10271Delta Opioid Receptor P ICC Rb Mm, Rt ab10272Kappa Opioid Receptor P ICC, IHC-Fr Rb Hu ab10283Kappa Opioid Receptor P WB Rb Mm ab10566Mu Opioid Receptor P IHC-P Rb Hu ab13074Mu Opioid Receptor P ICC, IHC-Fr, WB Rb Hu, Mm, Rt, Mk ab10275Mu Opioid Receptor P IP Rb Mm ab10582Mu Opioid Receptor splice variant P IHC-Fr Rb Mm, Rt ab10306Mu Opioid Receptor splice variant P ICC, IHC-Fr Gp Mm, Rt ab10273Mu Opioid Receptor peptide ab10704

Neuroscience > Neurotransmission > Receptors / Channels > GPCR > Serotonin Receptors


5HT1B Receptor P WB Rb Hu, Mm, Rt ab138965-HT1B Receptor P IHC-P Rb Hu ab131545HT1D Receptor P WB Rb Hu, Mm ab101325HT1D Receptor P WB Rb Hu, Mm, Rt ab138955-HT1E Receptor P IHC-P Rb Hu ab131555HT1F Receptor P IHC-P Rb Hu ab131565HT2B Receptor P IHC-P Rb Hu, Rt ab132925HT3 Receptor P IHC Rb Mm, Rt ab167885HT3 Receptor P WB Rb Hu, Rt ab138975HT4 Receptor P WB Rb Hu, Mm ab100955HT6 Receptor P IHC-P Rb Hu ab132935HT7 Receptor P IHC-Fr, IHC-P Rb Hu ab132945HT7 Receptor P WB Rb Hu, Mm, Rt ab13898Ketanserin M ELISA Mm Fast track ab1061

5HT2A Receptor antibody (ab16028)

IHC-FrFl - ab16028 staining 5HT2AR on rat brain tissue. Staining seen in coronal section through hypothalamus.Nuclear staining was observed in the Arcuate nucleus. Nuclear-like staining in the nucleus of the solitary tract, but notin the Ventromedial nucleus of the hypothalamus.

Clonality Applications tested Host Species reactivity DatasheetP IHC-FrFl, WB Rb Rt www.abcam.com/ab16028

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Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > AMPA / Kainate


Glutamate Receptor 1 (AMPA subtype)(phospho S831) P WB Rb Mm, Rt ab3902Glutamate Receptor 1 (AMPA subtype)(phospho T840) P IP, WB Rb Mm ab12108Glutamate Receptor 1 (AMPA subtype)(phospho S845) P WB Rb Mm, Rt ab3901KA1 P WB Rb Hu, Mm ab10101

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > GABA Receptors


GABA A Receptor alpha 1 P IP, WB Rb Rt ab5781GABA A Receptor alpha 2 P WB Rb Mm, Rt ab8342GABA A Receptor alpha 4 P IP, WB Rb Mm, Rt ab4120GABA A Receptor alpha 4 P Dot, WB Rb Rt ab13872GABA A Receptor alpha 5 P WB Rb Rt, Mm ab3700GABA A Receptor alpha 5 P WB Rb Hu, Mm ab10098GABA A Receptor alpha 6 P IP, WB Rb Mm, Rt ab5365GABA A Receptor beta 1 P IP, WB Rb Mm, Rt ab16703GABA A Receptor beta 1 P WB Rb Hu ab23338GABA A Receptor beta 2 P IP, WB Rb Mm, Rt ab8340GABA A Receptor beta 3 P IP, WB Rb Mm, Rt ab4046GABA A Receptor delta P WB Rb Hu, Mm ab10100 Pep. avail.GABA A Receptor gamma 2 P IP, WB Rb Mm, Rt ab4073GABA A Receptor gamma 3 P WB Rb Hu, Mm, Rt ab13861 Pep. avail.GABA B Receptor 1 P WB Rb Hu ab1135 Pep. avail.GABA B Receptor 2 P WB Rb Hu ab1134GABA B Receptor 2 (phospho S892) P IF, IHC, WB Rb Mm, Rt ab3900GABARAP P WB Rb Hu ab1133

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > Glycine Receptor


Glycine Receptor alpha 1 P IP, WB Rb Hu ab475Glycine Receptor alpha 1 + alpha 2 P IHC-Fr, WB Rb Rt ab23809 Pep. avail.

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > More Channels


alpha 1 Glycine Receptor P IP, WB Rb ab475

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > nAch Receptors


Nicotinic Acetylcholine Receptor alpha [D6] M FACS, IHC-Fr, IP, WB Mm Hu, Mm ab11149Nicotinic Acetylcholine Receptor [88B] M IHC-Fr, IP, WB Mm Rt, Mm, Ch, Fi, Am ab2804Nicotinic Acetylcholine Receptor beta [B3] M FACS, IHC-Fr, IP, WB Mm Hu.

Does not react with Mm, Rt or Ch ab11150

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Nicotinic Acetylcholine Receptor gamma [C9] M FACS, IHC-Fr, IP, WB Mm Foetal AChR.Does not react with Adult Hu,Mm, Rt or Ch ab11151

Rapsyn [1234] M ICC, IF, IHC-Fr, WB Mm Mm, Rt, Am, Ch, Fi ab11423 Pep. avail.Syntrophin [1351] M IF, IP, WB Mm Hu, Mm, Rt, Am, Ch, D, Fi ab11425Syntrophin gamma 2 P ELISA, WB Gt Hu ab15713

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > NMDA Receptors


NMDAR1 [R1JHL ] M WB Mm Rt ab1880NMDAR1 C1 P IHC, IP, WB Rb Rt ab6484NMDAR1 a/b/c/d P IHC, IP, WB Rb Rt ab6485NMDAR1 e/f/g/h P IHC, IP, WB Rb Rt ab6486NMDAR1 N1 P IHC, IP, WB Rb Rt ab6483NMDAR2A P IHC, IP, WB Rb Mm, Rt, Mk ab14596NMDAR2B P IP, WB Rb Mm ab14400NMDAR2B (phospho Y1336) P WB Rb Hu, Rt ab14965NMDAR2B (phospho Y1472) P Dot, WB Rb Hu, Mm, Rt ab16409NMDAR2C P IHC, IP, WB Rb Hu, Mm, Rt ab110NMDAR3A + 3B M WB Mm Hu ab2639

Neuroscience > Neurotransmission > Receptors / Channels > Ligand-Gated Ion Channels > P2X Receptors


P2X1 P ICC, IHC-Fr Rb Rt ab10248P2X2 P ICC, IHC-Fr, WB Gp Hu, Rt ab10262P2X2 P ICC, IHC-Fr, WB Rb Hu, Rt, Mk ab10266P2X3 P ICC, IHC-Fr, WB Rb Hu, Rt, Mk ab10269

Neuroscience > Neurotransmission > Receptors / Channels > More Ion Channels


ARA9 [35-2] M WB Mm Hu, Mm ab468ASIC beta P IHC-Fr Gp Rt ab10355DDR1 P ELISA, WB Rb Hu ab5508DDR2 P ELISA, WB Rb Hu ab5520ENSA P WB Ch Hu, Mm, Rt, B ab14297GJB1 P IHC-Fr, WB Rb Hu, Rt ab11367GJB1 [CXN-32] M ELISA, IHC-Fr, WB Mm Hu, Mm, Rt ab11366SLC31A1 P IHC-P Rb Hu ab13828TRAR4 P IHC-P Rb Hu ab13139TRPM7 P WB Gt Hu ab729TRPM7 P IP Rb Hu.

Does not react with Mm or Rt ab16576Vanilloid Receptor 1 P IF, IHC-Fr, IHC-P Rb Rt ab6166Vanilloid Receptor 1 P ICC, IHC-Fr Rb Rt ab3486Vanilloid Receptor 1 P IHC-Fr Gp Rt ab10295 Pep. avail.Vanilloid Receptor 1 P ICC, IHC-Fr, WB Rb Mm, Rt, Mk ab10296 Pep. avail.VRL1 P Dot, IHC-Fr, WB Rb Rt ab6183

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Neuroscience > Neurotransmission > Receptors / Channels > Potassium Channels


HCN1 P WB Rb Rt ab19405KChIP2 P WB Rb Hu, Rt ab3473KCNQ3 P ICC, IF, IHC-Fr Rb Hu, Mm, Rt ab16228KCNQ5 P WB Rb Rt ab19319 Pep. avail.Kv beta 2 P ICC, WB Rb Mm ab10665Kv12 P ICC, WB Rb Mm, Rt ab10667Kv12 [0T76] M IHC, IP, WB Mm Mm, Rt, Mk ab21091Kv14 P IF, IP, WB Rb Hu ab16718Kv22 - Azide free P IHC, WB Rb Rt, X ab21909Kv42 P IF, IP, WB Rb Rt ab16719

Neuroscience > Neurotransmission > Receptors / Channels > Sodium Channels


APXL P Fast track Gt Fast track ab5948ASIC3 P ICC, IHC-Fr Gp Rt ab10354

Neuroscience > Neurotransmission > Receptors / Channels > Tyrosine Kinase Receptors


EphA1 P ELISA, WB Rb Hu ab5385EphA1 P I-ELISA, WB Gt Hu ab7036EphA2 P ELISA, WB Rb Ha, Hu, Mm ab5387EphA2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610EphA3 P I-ELISA, WB Gt Mm ab7038EphA3 P ELISA, WB Gt Hu. Does not react with Mm ab10611EphA3 [III-A4] M FACS, IP Mm Hu ab19439EphA4 P ELISA, WB Rb Hu ab5389EphA4 P I-ELISA, WB Gt Mm ab7039EphA4 P I-ELISA, WB Gt Hu ab7041EphA5 P ELISA, WB Rb Hu, Mm ab5397EphA5 P ELISA, WB Gt Mm, Rt ab10612EphA6 P ELISA, WB Gt Mm ab10613EphA7 P ELISA, WB Rb Hu ab5411EphA7 P ELISA, WB Gt Mm ab10614EphA8 P ELISA, WB Gt Mm ab10615EphB1 P ELISA, WB Rb Hu, Mm ab5414EphB1 P I-ELISA, WB Gt Rt ab7042EphB1 P Dot, IP Sh Hu ab10406EphB2 P ELISA, WB Rb Hu, Mm ab5418EphB2 P Dot, ELISA Gt Mm ab10616EphB3 P ELISA, WB Gt Mm ab10617EphB3 P ELISA, WB Gt Hu ab7044EphB4 P Dot, ELISA, Inhib Gt Mm ab10618EphB6 P Dot, ELISA Gt Mm ab10619Ephrin A2 P BL, ELISA, ICC, IHC-Fr, WB Gt Mm ab10610Ephrin A3 P ELISA, WB Gt Hu. Does not react with Mm ab10611Ephrin A4 P I-ELISA, WB Gt Hu ab7041Ephrin B3 P ELISA, WB Gt Hu ab7044

Neuroscience > Neurotransmission > Secretory Vesicles > Granins


Secretogranin II P IHC, IP, WB Rb Hu, Rt ab12241

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Neuroscience > Neurotransmission > Secretory Vesicles > Munc18


Mint1 P WB Rb Rt ab3448Munc18 P IP, WB Rb Rt ab3451MYRIP P Fast track Gt Fast track ab10149Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260Synaptotagmin P IF, WB Rb Hu, Rt ab10104Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259

Neuroscience > Neurotransmission > Secretory Vesicles > Related targets


Amisyn P Fast track Gt Fast track ab5899Calpastatin P ELISA, WB Ch Hu ab16423Calpastatin [2G11D6] M ICC, WB Mm Hu, Rt, C, Pi ab3515Clathrin light chain [2Q2211] M WB Mm B ab16428Clathrin light chain [3F133] M IF, IHC-P, IP, WB Mm C ab14408Clathrin light chain [3F133] - BSA and Azide free M ICC, IF, IHC-P, IP, WB Mm Hu, C ab14409 Pep. avail.Clathrin light chain [CON1] M AP, ICC, IHC, WB Mm Hu, Mm, Rt, C, Sc ab11332 Pep. avail.Clathrin light chain [SPM174], prediluted M IHC-P Mm Hu, C ab17949Mint1 P WB Rb Rt ab3448Mint3 P WB Rb Rt, Mm ab3450MYRIP P Fast track Gt Fast track ab10149SCAMP1 P IP, WB Rb Rt, Pi ab3430SCAMP2 P WB Rb Hu, Mm, Ha ab3431SCAMP5 P WB Rb Rt ab3432Secretogranin II P IHC, IP, WB Rb Hu, Rt ab12241SLC6A4 [4A22] M FACS, IHC-P Mm Hu, Rt ab1125Synaptotagmin P ICC, IP, WB Rb Rt, X ab13260Synaptotagmin P IF, WB Rb Hu, Rt ab10104Synaptotagmin P ELISA, WB Ch Hu, B, Mm, Pi, Rt ab8037Synaptotagmin [ASV30] M ICC, IHC, IP, WB Mm Mm, Rt, X, B, Ch, Rb, Sha ab13259Synaptotagmin [ASV48] M ICC, IHC, IP, WB Mm Rt, B, Mk, Sha, Mm ab12255

Neuroscience > Neurotransmission > Secretory Vesicles > Rabs


Rab5 P ICC, IHC, IP, WB Rb Hu, Mm, Rt, B, D, Gp, Ha, Mk ab13253Rabphilin 3A P WB Rb Rt ab3338Rabphilin 3A P WB Rb Mm, Rt, B ab12234

Neuroscience > Neurotransmission > Secretory Vesicles > SNAPs & SNAREs


Amisyn P Fast track Gt Fast track ab5899SNAP23 P IF, WB Rb Hu, Rt ab4114SNAP23 P WB Rb Rt, Mm ab3340SNAP25 P IF, WB Rb Mm, Rt, B ab5666 Pep. avail.SNAP25 [4H251] M WB Mm Hu, Mm, Rt, X, B, Ha, In (tick), Pi ab18002 Pep. avail.SNAP25 [SP12] M ELISA, IHC-P, WB Mm Hu, Ha, Rt, Pi ab11102Synapsin I (phospho S603) P Dot, WB Rb Rt ab13879Synapsin I (phospho S9) P WB Rb Hu, Mm, Rt, X , B ab16554

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Synapsin I (phospho S9) P WB Rb Hu ab18006Synapsin I P AF, IF, IHC-F, IHC-Fr, IP, WB Rb B, Gp, Hu, Mm, Rt ab8 Pep. avail.Synapsin I P IP, WB Rb Hu, Mm, Rt, B ab12239 Pep. avail.Synapsin I P WB Rb Hu, Rt ab18004Synaptobrevin [4E240] M WB Mm Hu, Mm, Rt, Rb, X, B, Gp,

Ha, Mk, Pi, Sh ab18013Synaptobrevin [4H302] M WB Mm Hu, Mm, Rt, X, B, Ha, Pi ab18015Synaptobrevin [SP10] M WB Mm Hu, Rt, Ha, Pi ab11109Synaptobrevin [SP11] M ELISA, IHC-P, WB Mm Hu, Pi.

Does not react with Rt ab11104Synaptophysin P IHC-F, IHC-Fr, IHC-P Rb Hu ab7837Synaptophysin P IHC-Fr, IP, WB Rb Hu, Mm, Rt ab14692Synaptophysin [4E206] M IHC, IP, WB Mm Hu, Mm, Rt, B ab18008Synaptophysin [4H255] M WB Mm Hu, Mm, Rt, B ab18007Synaptophysin [SP11] M IHC-P, WB Rb Hu ab16659Synaptophysin [SP15] M ELISA, IHC-P, WB Mm Hu, Rt, Ha, Pi ab11105Synaptophysin [SVP38] M FACS, ICC, WB Mm Rt ab12353Synaptophysin [SY38] M IHC-Fr, IHC-P Mm Rt, Mm, B ab8049Synaptophysin [SYP02] M IHC-Fr, IHC-P Mm Hu ab6245Synaptophysin, prediluted P IHC-F, IHC-P Rb Hu ab8547 Pep. avail.Syntaxin 3 P IF, WB Rb Ma ab4113Syntaxin P ELISA, IP, WB Ch Hu, B, Mm, Pi, Rt ab8038Syntaxin P WB Rb Mm, Rt, C ab13262Syntaxin [1B663] M IP, WB Mm Rt ab18009Syntaxin [4H256] M WB Mm Hu, Mm, Rt, X, B, F, Ha, Pi ab18010Syntaxin [SP6] M IHC-Fr, IP, WB Mm Hu, Mm, Rt, X, B, F, Ha, Pi ab12236Syntaxin [SP8] M ELISA, IHC-Fr, WB Mm Hu, Rt, Pi, Ha ab257Syntaxin [STX01 (HPC-1)] M IHC-P, WB Mm Hu, Rb, Rt, C.

Does not react with Gp ab3265VAMP8 P IF, IM, IP, WB Rb Hu, Rt, D ab6186

Neuroscience > Neurotransmission > Transporters > Dopamine


CSPS P WB Rb Hu, Mm, Rt ab16870CSPS [3F10] M ELISA, IP Mm Hu, Mm, Rt ab16773Dopamine Transporter P ELISA, WB Rb Hu, Mm, Rt ab18548Dopamine Transporter [hDAT-LOOP] M ICC, IHC-Fr, WB Rt Hu, Rt ab5981

Neuroscience > Neurotransmission > Transporters > GABA


GABA Transporter 1 / GAT 1 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab426GABA Transporter 2 P IF, IHC-Fr, WB Rb Hu, Mm, Rt, Ch ab2896GABA Transporter 3 / GAT 3 P IF, IHC-Fr, IHC-P, WB Rb Hu, Mm, Rt, Ch ab431

Neuroscience > Neurotransmission > Transporters > Glutamate


Glutamate Aspartate Transporter P FACS, ICC, IHC, PCC, WB Rb Rt ab416Glutamate Transporter 1 P FACS, ICC, IF, IHC, PCC, WB Rb Hu, Rt ab417VGLUT1 P EM, IHC, IP, RIA, WB Rb Hu, Rt ab18106

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Neuroscience > Sensory systems > Hearing


MATH1 P ELISA, IF, IHC-Fr, WB Rb Hu, Mm, Rt ab13483TBL1 P WB Gt Hu ab2243 Pep. avail.

Neuroscience > Sensory systems > Nociception


5 Methoxytryptamine P IHC Rb Rt ab15165Amitriptyline [1BB831] M ELISA, RIA Mm Fast track ab15176Amitriptyline [202] - BSA and Azide free M IA Mm Fast track ab20408ASIC beta P IHC-Fr Gp Rt ab10355ASIC3 P ICC, IHC-Fr Gp Rt ab10354Axotrophin P WB Ch Hu, Mm, Rt ab13997Clonidine P IP Rb Many ab14410Endorphin alpha neo P IHC-Fr, RIA Rb Rt, Pi ab11140Neuropeptide FF1 P WB Rb Hu ab3898NMUR1 P IHC-P Rb Hu ab13365NMUR2 P IHC-P Rb Hu ab13366Nociceptin P IHC-Fr Gp Rt, Mk ab10276Nociceptin P IHC-Fr Rb Rt, Mk ab10277Nociceptin P ELISA, I-ELISA, IHC-Fr, WB Rb Gp, Rt ab6174NPFF1 Receptor P IHC-P Rb Hu ab13367NPFF1 Receptor P ELISA, WB Rb Hu ab1401NPFF2 P WB Rb Hu ab3899NPFF2 Receptor P IHC-P Rb Hu ab13198NPFF2 Receptor P ELISA, WB Rb Hu ab1402PSD93 P IHC-Fr, WB Rb Rt ab2930PSD93 P IHC-P, WB Gt Mm ab12097TRPM8 P WB Rb Hu ab3243Vanilloid Receptor 1 P IHC-Fr Gp Rt ab10295Vanilloid Receptor 1 P ICC, IHC-Fr, WB Rb Mm, Rt, Mk ab10296Vanilloid Receptor 1 P IF, IHC-Fr Rb Hu ab3487

SPRR1a antibody (ab18580)

IF - detected in mouse L4 DRG sciatic nerve crush using ab18580 at 1/1000

Clonality Applications tested Host Species reactivity DatasheetP WB, IF Rb Mm. Does not react with Rt www.abcam.com/ab18580

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Vanilloid Receptor 1 P IF, IHC-Fr, IHC-P Rb Rt ab6166VRL1 P Dot, IHC-Fr, WB Rb Rt ab6183

Neuroscience > Sensory systems > Olfaction


Olfactory Receptor LS189744 P IHC-P Rb Hu ab13071Olfactory Receptor OR10R2 P IHC-P Rb Hu ab13203Olfactory Receptor OR2A4 P IHC-P Rb Hu ab13205Olfactory Receptor OR6N1 P IHC-P Rb Hu ab13204OR10R2 P IHC-P Rb Hu ab13072OR2H3 P IHC-P Rb Hu ab13073

Neuroscience > Sensory systems > Touch


OA1 P IHC-P Rb Hu ab13070

Neuroscience > Sensory systems > Vision


APXL P Fast track Gt Fast track ab5948Arrestin C P WB Ch Hu ab15852Bestrophin P ICC, IHC, IP, WB Rb Hu, Mm, Rt ab14927Bestrophin M WB Mm Hu, Pi, Mk ab2182CHX10 P WB Sh Hu, Mm, Rt, B ab16141CHX10 P WB Sh Hu, Mm, Rt, B ab16142Melanopsin P IF, IHC-P, WB Rb Mm, Rt ab19306Melanopsin P IF, IHC-Fr, IHC-P, WB Rb Rt. Does not cross-react with Mm ab19383NCKX2 P IHC-Fr, WB Rb Rt ab3507OA1 P IHC-P Rb Hu ab13070PDE6 alpha P WB Rb C, Sh ab5659PDE6 alpha P WB Rb Sh ab5660Peropsin P IHC-P Rb Hu ab13377RAGE P IHC-Fr, WB Rb Rt, Mm ab3611RAGE P ELISA, IHC-P Gt Hu ab20558RIT1 P WB Rb Hu ab13322RIT1 [14G7] M WB Mm Hu, Mm ab16458

Vanilloid Receptor 1 antibody (ab3486)

ICC - stained cells in dorsal root ganglion cells. ab3486 (1/10,000 for 2h at RT) on adult rat dorsal ganglion cells(cultured for 48hrs and fixed in 4% PFA/15% picric acid).

Clonality Applications tested Host Species reactivity DatasheetP IHC-Fr, ICC Rb Rt www.abcam.com/ab3486

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Product datasheet guide


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CONTENTS Page

I. WESTERN BLOTTING (WB) – A BEGINNER’S GUIDE. . . . . . . . . . . 66A. Lysate preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67B. Preparation of cell lysate from cells in culture . . . . . . . . . . . . . . . . 67C Preparation of cell lysate form tissues . . . . . . . . . . . . . . . . . . . . . . 67D. Determination of the protein concentration levels . . . . . . . . . . . . . 67E. Denaturation and reduction of the protein . . . . . . . . . . . . . . . . . . . 68F. Electrophoresis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68G. Transfer of proteins onto a membrane . . . . . . . . . . . . . . . . . . . . . . 69H. Blocking the membrane . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69I. Incubation with primary antibody . . . . . . . . . . . . . . . . . . . . . . . . . . 70J. Incubation with secondary antibody . . . . . . . . . . . . . . . . . . . . . . . . 70K. Development methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70L. Determination of the protein size . . . . . . . . . . . . . . . . . . . . . . . . . . 70M. Western blot protocol troubleshooting tips . . . . . . . . . . . . . . . . . . . 70N. Stripping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

II. WESTERN BLOTTING OF PHOSPHO-PROTEINS PROTOCOL . . . 72

III. IMMUNOPRECIPITATION PROTOCOL (IP) . . . . . . . . . . . . . . . . . . . 73A. Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73B. Immunoprecipitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73

IV. NUCLEAR FRACTIONATION PROTOCOL . . . . . . . . . . . . . . . . . . . 73A. Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73B. Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74

V. MITOCHONDRIAL PURIFICATION PROTOCOL. . . . . . . . . . . . . . . . 74

VI. SOLUBLE (S-100) MITOCHONDRIAL FRACTIONATION . . . . . . . . 74PROTOCOL

VII. DETERMINING IF THE ANTIBODY BINDS ONLY . . . . . . . . . . . . . 74PHOSPHORYLATED PROTEINS (WB OR IHC)

VIII. DOT BLOT PROTOCOL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74A. Reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74B. Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74

IX. BLOCKING WITH IMMUNIZING PEPTIDE (BL) PROTOCOL . . . . . 75

X. IMMUNOHISTOCHEMISTRY (IHC-P) – PARAFFIN PROTOCOLS. . . 75A. Antigen retrieval . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75B. ABC immunohistochemical staining – chromogenic methods . . . . 76C. ABC immunohistochemical staining – fluorescent methods . . . . . 78D. IHC troubleshooting tips . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

XI. IMMUNOHISTOCHEMISTRY (IHC-FR) - FROZEN SECTIONS . . . 79

XII. IMMUNOCYTOCHEMISTRY (ICC) PROTOCOL . . . . . . . . . . . . . . . 79

XIII. DOUBLE IMMUNOFLUORESCENCE (IF) STAINING . . . . . . . . . . 80– SIMULTANEOUS PROTOCOL

XIV. DOUBLE IMMUNOFLUORESCENCE (IF) STAINING . . . . . . . . . . 80– SEQUENTIAL PROTOCOL

XV. INDIRECT FLOW CYTOMETRY (FACS) PROTOCOL . . . . . . . . . . 81

XVI. DIRECT FLOW CYTOMETRY (FACS) PROTOCOL . . . . . . . . . . . 81

XVII. DIRECT ELISA PROTOCOL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82

XVIII. INDIRECT ELISA PROTOCOL . . . . . . . . . . . . . . . . . . . . . . . . . . . 82

XIX. ELISA USING FLUORESCENT SUBSTRATE . . . . . . . . . . . . . . . . 83

XX. CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL . . . . 83

XXI. BUFFERS AND STOCK SOLUTIONS . . . . . . . . . . . . . . . . . . . . . . 83

I. WESTERN BLOTTING (WB) - A BEGINNER’S GUIDE

Western blotting (or immunoblotting) enables determination of the relativeamounts of a specific protein present in different samples by separatingproteins on a gel and probing for the specific protein with an antibodytargeted to that protein only.

Here is a beginner’s guide to western blotting and our recommendationswhen using Abcam antibodies.

A) Lysate preparation

Lysis buffersA number of lysis buffers are available and the choice of lysis bufferdepends on the protein investigated. Where the target protein is tightlybound to cytoskeletal elements, a stronger extraction buffer containingSDS or other detergents is required (see below).

Cytoplasmic protein: Use NP-40 or RIPA buffer.Soluble proteins are the easiest proteins to extract from a cellhomogenate; thus, this can be done readily even without detergent.

Membrane-bound protein: Use NP-40 or RIPA buffer.Transmembrane proteins are contained within membranes, a suitabledetergent is essential to extract these proteins.

Nuclear protein: Use RIPA buffer. (see page 67)

Mitochondrial protein: Use Lysis buffer. (see page 74)

Whole cell protein extracts: Use NP-40 or RIPA buffer.

In summary...

Cytoskeletal bound proteins Extract Buffer:10 mM Tris, pH 7.4100 mM NaCl1 mM EDTA1 mM EGTA1 mM NaF20 mM Na4P2O7

2 mM Na3VO4

1% Triton X-10010% glycerol0.1% SDS0.5% deoxycholate

Soluble protein buffer:20 mM Tris-HCl, pH 7.51 mM EGTA (Ca2+ chelator)

If in doubt, start by using RIPA buffer as it is the “strongest”.

Protein location Buffer recommended

Cytoplasmic (soluble) Tris buffer

Cytoplasmic (cytoskeletal bound) Tris-Triton buffer

Membrane bound NP-40 or RIPA buffer

Nuclear RIPA buffer

Mitochondrial Use specific protocol

Whole Cell NP-40 or RIPA buffer

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RIPA buffer (RadioImmunoPrecipitation Assay) buffer:RIPA buffer contains the ionic detergent sodium deoxycholate as an activeconstituent and is particularly used for nuclear membrane disruption fornuclear extracts. RIPA buffer gives low background but can denaturekinases. It can also disrupt protein-protein interactions (and may thereforebe problematic for immunoprecipitations/pull down assays).

50mM Tris HCl pH 8150 mM NaCl1% NP-40 0.5% sodium Deoxycholate 0.1% SDS

The 10% sodium deoxycholate stock solution (5 g into 50 ml) must beprotected from light.The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O andthen add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the totalvolume to 50 ml). Store the buffer at 4°C.

Nonidet-P40 (NP-40) buffer:20 mM Tris HCl pH 8137 mM NaCl10% glycerol1% nonidet P-402 mM EDTA

Protease inhibitorsAs soon as lysis occurs, proteolysis, dephosphorylation and denaturationbegin. These events can be slowed down tremendously if samples arekept on ice or at 4°C at all times and appropriate inhibitors are addedfresh to the lysis buffer.

EDTA chelated calcium and magnesium ions.Sodium orthovanadate is an inhibitor of all tyrosine protein phosphatases.It must be prepared correctly (see below).Sodium fluoride is an inhibitor of serine/threonine protein phosphatases.Aprotinin is a protease inhibitor.Leupeptin is a lysosomal protease inhibitor.Anti-pain inhibits trypsin and plasmin proteases.Pepstatin is a reversible inhibitor of aspartic proteases.PMSF is a protease inhibitor.

Ready to use cocktails of inhibitors from well known suppliers are oftenused but you can make your own cocktail.

Sodium orthovanadate preparation:This needs to be done under the fume hood

• Prepare a 100 mM solution in double distilled water• Set pH to 9.0 with HCl• Boil until colorless• Cool to room temperature • Set pH to 9.0 again• Boil again until colorless• Repeat this cycle until the solution remains at pH 9.0 after boiling and

cooling• Bring up to the initial volume with water• Store in aliquots at - 20°C

Note: do not permit great changes in volume during boiling; put a loose lidon the container to protect from evaporation.Discard if the samples turn yellow.

Remember to add phosphatase inhibitors to cocktailsbought when investigating phosphorylation events.

Final concentrations of inhibitors to use:

The buffer (with inhibitors) should be ice-cold prior tohomogenization.

B) Preparation of cell lysate from cells in culturePlace the cell culture dish in ice and wash the cells with ice-cold PBS,drain the PBS, then add ice-cold lysis buffer (1ml per 107 cells/100mmdish/150cm2 flask; 0.5ml per 5x106 cells/60mm dish/75cm2 flask). Scrapeadherent cells off the dish using a cold plastic cell scraper then gentlytransfer the cell suspension into a pre-cooled microfuge tube.

Maintain constant agitation for 30 minutes at 4°C.

Centrifuge in a microcentrifuge at 4°C. You may have to vary thecentrifugation force and time depending on the cell type, a guideline is 20min at 12000 rpm but this must be determined by the end-user (e.g.leukocytes need a very light centrifugation).

Gently remove the tubes from the centrifuge and place on ice, aspirate thesupernatant and place in a fresh tube kept on ice, and discard the pellet.

C) Preparation of cell lysate from tissuesDissect the tissue of interest with clean tools, on ice preferably, and asquickly as possible to prevent degradation by proteases. Place the tissuein round bottom microfuge tubes and immerse in liquid nitrogen to “snapfreeze”. Store samples at -80°C for later use or keep on ice for immediatehomogenization.

For a ~5 mg piece of tissue, add ~300 µl lysis buffer rapidly to the tube,homogenize with an electric homogenizer, rinse the blade twice withanother 2x300 µl lysis buffer, then maintain constant agitation for 2 hoursat 4°C (e.g place on an orbital shaker in the fridge). Volumes of lysis buffermust be determined in relation to the amount of tissue present (proteinextract should not be too diluted to avoid loss of protein and large volumesof samples to be loaded onto gels, the minimum concentration is 0.1mg/ml, optimal concentration is 1-5 mg/ml).

Centrifuge for 20 min at 12000 rpm at 4°C in a microcentrifuge. Gentlyremove the tubes from the centrifuge and place in ice, aspirate thesupernatant and place in a fresh tube kept on ice, discard the pellet.

D) Determination of the protein concentration levelsPerform a Bradford assay, a Lowry assay or a BCA assay. Bovine serumalbumin (BSA) is a frequently used protein standard.

Inhibitor Proteaseinhibited

Finalconcentration in

lysis buffer

Stock(store at -20°C)

AprotininTrypin,

chynotrypin,plasmin

2 µg/ml Dilute in water.Do not re-use

when defrosted.

Leupeptin Lysosomal 5-10 µg/mlDilute in water.Do not re-use

when defrosted.

Antipain Trypin, plasmin 2-10 µg/mlDilute in water.Do not re-use

when defrosted.

Pepstatin A aspartyl 1 µg/mlDilute in water.Do not re-use

when defrosted.

Na-Fluoride Serine/Threoninephosphatase 5-10 mM

Dilute in water.Do not re-use

when defrosted.

Orthovanadate Tyrosinephosphatase 1 mM

Dilute in water.Do not re-use

when defrosted.

PMSF Serine, cysteine 1 mMDilute in ethanol,you can re-use

the same aliquot.

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Once you have determined the concentration in each sample, you canthen freeze them at -20°C or -80°C for later use or proceed to theimmunoprecipitation stage (see page 73).

E) Denaturation and reduction of the proteinAntibodies typically recognize a small portion of the protein of interest(termed epitope) and this domain may reside within the 3D conformation ofthe protein. To enable access of the antibody to this portion it is necessaryto unfold the protein, i.e. denature it.

To do so, use a loading buffer with the anionic denaturing detergent sodiumdodecyl sulfate (SDS), and boil the mixture at 95-100°C for 5 minutes.

When SDS is used with proteins, all of the proteins become negativelycharged by their attachment to the SDS anions. SDS denatures proteinsby “wrapping around” the polypeptide backbone - and SDS binds toproteins fairly specifically in a mass ratio of 1.4:1. In so doing, SDSconfers a negative charge to the polypeptide in proportion to its length -i.e., the denatured polypeptides become “rods” of negative charge cloudswith equal charge or charge densities per unit length. It is usuallynecessary to reduce disulphide bridges in proteins before they adopt therandom-coil configuration necessary for separation by size by using ß-mercaptoethanol or dithiothreitol (DTT). In denaturing SDS-PAGEseparations, therefore, migration is determined not by intrinsic electricalcharge of the polypeptide, but by molecular weight. SDS grade is of utmost importance: a protein stained background alongindividual gel tracts with indistinct or slightly distinct protein bands areindicative of old or poor quality SDS.

Exceptions:

1) Certain antibodies recognize the native form (i.e. non-denatured form)of the protein. It is imperative in those circumstances to run a Western Blotin non-denaturing conditions.

2) Certain antibodies only recognize protein in its non-reduced form i.e. inan oxidized form (particularly on cysteine residues) and the compoundSDS must be removed from the traditional loading buffer and migrationbuffer (non reducing conditions).

In summary…

Rule of thumb: Use a reducing and denaturing WBcondition unless the datasheet specifies otherwise.

To enable visualization of the migration of proteins it is common to includein the loading buffer a small anionic dye molecule (e.g., bromophenolblue). Since the dye is anionic, and small, it will migrate the fastest of anycomponent in the mixture to be separated and provide a migration front tomonitor the separation progress.Glycerol is also added to the loading buffer to increase the viscosity of thesample to be loaded and hence maintain the sample at the bottom of thewell, restricting overflow and uneven gel loading.During protein sample treatment the sample should be mixed by vortexingbefore and after the heating step for best resolution.

F) ElectrophoresisElectrophoresis can be one dimensional (i.e., one plane of separation) ortwo dimensional. One dimensional electrophoresis is used for most routineprotein and nucleic acid separations. Two dimensional separation of

proteins is used for finger-printing, and when properly constructed can beextremely accurate in resolving all of the proteins present within a cell(greater than 1,500 KDa).

Here we will be describing techniques for one dimensional electrophoresis.We recommend Gel Electrophoresis, A Practical Approach (2nd Edition, BDHames and D Rickwood, The Practical Approach Series, IRL press ) as areference for basic understanding of 2D electrophoresis protocols.

i) Preparation of PAGE gelsWhen separated on a polyacrylamide gel, the procedure is abbreviated asSDS-PAGE (for Sodium Dodecyl Sulfate PolyAcrylamide GelElectrophoresis). The technique has become a standard means formolecular weight determination.

Polyacrylamide gels are formed from the polymerization of twocompounds, acrylamide and N,N-methylene- bis-acrylamide (Bis, forshort). Bis is a cross-linking agent for the gels. The polymerization isinitiated by the addition of ammonium persulfate along with either DMAPor TEMED. The gels are neutral, hydrophilic, three-dimensional networksof long hydrocarbons crosslinked by methylene groups. The separation of molecules within a gel is determined by the relative sizeof the pores formed within the gel. The pore size of a gel is determined bytwo factors, the total amount of acrylamide present (designated as%T) andthe amount of cross-linker (%C). As the total amount of acrylamideincreases, the pore size decreases. With cross- linking, 5%C gives thesmallest pore size. Any increase or decrease in%C increases the poresize. Gels are designated as percent solutions and will have twonecessary parameters. The total acrylamide is given as a% (w/v) of theacrylamide plus the bis-acrylamide. Thus, a 7.5%T would indicate thatthere is a total of 7.5 g of acrylamide and bis per 100 ml of gel. A geldesignated as 7.5%T:5%C would have a total of 7.5% (w/v) acrylamide +bis, and the bis would be 5% of the total (with pure acrylamide composingthe remaining 2.5%).Gels can be purchased ready-made (the Abcam laboratory uses Nu-PageTM gels from Invitrogen) or tailor-made in the laboratory (recipes canbe found in laboratory handbooks). Either way, choose carefully thepercentage of your gel as this will determine the rate of migration anddegree of separation between proteins.

Rule of thumb: The smaller the size of the protein of interest, the higherthe percentage of mono/bis.The bigger the size of the protein of interest, the lowerthe percentage of mono/bis.

Acrylamide is a potent cumulative neurotoxin: wear gloves at alltimes.

Place gels in the electrophoresis tank as instructed by the manufacturerand bathe in migration buffer.

ii) Use of positive controlsA positive control lysate is used to demonstrate that the protocol is efficientand correct and that the antibody recognizes the target protein which maynot be present in the experimental samples.

We strongly recommend the use of a positive controllysate when setting up a new experiment; this will giveyou immediate confidence in the protocol.

iii) Use of a molecular weight markerA range of molecular weight markers will enable the determination of theprotein size (see below) and also to monitor the progress of anelectrophoretic run. A range of MW markers are commercially available,

Protein size (kDa) Gel percentage (%)

4-40 20

12-45 15

10-70 12.5

15-100 10

25-200 8

Protein State WB condition Loading buffer Migration buffer

Reduced -Denatured

Reducing &Denaturing

With SDS &ß-mercaptoethanol

or DTTWith SDS

Reduced- Native Native = Non-Denaturing No SDS No SDS

Oxidized -Denatured Non-Reducing

With SDS, Noß-mercaptoethanol,

No DTTWith SDS

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we would recommend colored markers for convenience (i.e., each MWband size will be of a different color and therefore easily detectable).

iv) Loading the samples onto the gelUse special gel loading tips or a micro-syringe to load the completesample in a narrow well with limited movement at the bottom of the well,resulting in high quality data.Never overfill wells. This could lead to artefacts.Load 20-40 µg total protein per mini-gel well. The gels will be submerged in migration buffer which normally containsSDS, except in native gel electrophoresis.

Run the gel for the recommended time as instructed by the manufacturer,this can vary from machine to machine (1 hour to overnight).When the dye molecule (the “migration front”) reaches the bottom of thegel, the power is turned off and the experiment halted.

v) Loading control troubleshooting tipsLoading controls are required to check that the lanes in yourgel have been evenly loaded with sample, especially when acomparison must be made between the expression levels ofa protein in different samples.They are also useful to check for even transfer from the gel to themembrane across the whole gel. Where even loading or transfer have notoccurred, the loading control bands can be used to quantify the proteinamounts in each lane. For publication-quality work, use of a loadingcontrol is absolutely essential.

G) Transfer of proteins onto a membrane (WB)Transfer can be done in wet or semi-dry conditions. Wet conditions arerecommended; especially for high MW proteins (if a semi-dry transfersystem is used for large proteins add SDS to help transfer). Use the powerconditions recommended by the manufacturer.

Two types of membranes are available: nitrocellulose and PVDF. Thechoice is personal and both work very well.

PVDF membranes require careful pre-treatment: cut the membrane to theappropriate size then soak it in methanol for 1-2 min. Incubate in ice coldtransfer buffer for 5 min.

The gel needs to equilibrate for 3-5 min in ice cold transfer buffer. Failureto do so will cause shrinking while transferring and an unsightly pattern oftransfer.

i) Visualization of all the separated proteins after migrationThis visualization of protein at this stage is useful to determine if proteinshave migrated uniformly and evenly.

a) Coomassie stainAlthough the gel support provides some friction to molecular motion, assoon as the power is turned off the separated protein bands will begin todiffuse (they are freely soluble in aqueous solution). To prevent diffusionof proteins treat the gel with an acetic acid and methanol solution whichcauses almost all proteins to precipitate (become insoluble). To visualizethe fixed proteins use Coomassie Brilliant Blue R-250 (“Coomassie stain”)and then wash the stain out of the gel by incubation in a weak solution ofacetic acid and methanol. The stain will not bind to the acrylamide, andwill wash out (leaving a clear gel). However, it remains strongly bound tothe proteins in the gel, and these take on a deep blue color.

Please note: this is not reversible so if you choose to transfer the proteinsopt for a copper stain method to check migration is adequate.

b) Copper StainSoak freshly-electrophoresed gels in 0.3 M CuCl2 for up to 15 min,washing the gels briefly in de-ionized water, and view them against a dark-field background. Proteins come up as clear zones in a translucent bluebackground.

Gels may be destained completely by repeated washing in 0.1- 0.25 MTris/0.25 M EDTA pH 8.0, and then electroblotted, or eluted from the gelfor other purposes.

Visualization of protein at this stage is useful to determine if proteins havemigrated uniformly and evenly.

ii) Visualization after transfer onto membrane: Ponceau RedWash the membrane in TBST. Dilute the stock Ponceau Red 1:10. Thestock is made of 2% Ponceau S in 30% trichloroacetic acid and 30%sulfosalicylic acid. Incubate on an agitator for 5 min. Wash extensively inwater until the water is clear and the protein bands are well-defined.

The membrane may be destained completely by repeated washing inTBST or water. When using a PVDF membrane, re-humidify themembrane with methanol then wash again in incubation buffer (TBST).

TBST (also called TBS) is a Tris Buffered Saline Tween20 buffer. Somelaboratories use PBST but we recommend a tris based buffer.

TBS 10x (concentrated TBS)24.23 g Trizma HCl80.06 g NaClMix in 800 ml ultra pure water. pH to 7.6 with pure HCl.Top up to 1 L.

TBSTFor 1 L: 100 ml of TBS 10x + 900 ml ultra pure water + 1ml Tween20

Tween20 is very viscous and will stick to the tip of your measuring pipettesso make sure you add the right amount of the detergent to the tris buffer.

Keep TBST at 4°C to prevent contamination and do not keep it for morethan 1 week.

Do not add azide to TBST as it will quench the enzyme horse radishperoxidase (HRP) conjugated to the secondary antibody and preventdevelopment of the signal.

H) Blocking the membraneBlocking the membrane prevents non-specific background binding of theprimary and/or secondary antibodies to the membrane (which has a highcapacity at binding proteins and therefore antibodies). Two blocking solutions are traditionally used: non-fat milk or BSA.

Milk is cheaper but is not recommended for studies of phosphoproteins(milk contains casein which is a phosphoprotein; this is why it causes highbackground because the phospho-specific antibody detects the caseinpresent in the milk).

Sample type Loadingcontrol

Molecularweight (kD)

Caution

Whole cell/cytoplasmic

beta actin 43 Not suitable for skeletalmuscle samples. Changes incell-growth conditions andinteractions with extracellularmatrix components may alteractin protein synthesis (Farmeret al., 1983).

GAPDH 30-40 Some physiological factors,such as hypoxia and diabetes,increase GAPDH expression incertain cell types.

tubulin 55 Tubulin expression may varyaccording to resistance toantimicrobial and antimitoticdrugs (Sangrajrang S. et al,1998, Prasad V et al, 2000)

Mitochondrial VDAC1 /Porin

31

COXIV 16 Many proteins run at the same16 kD size as COXIV.

Nuclear Lamin B1 66 Not suitable for samples wherenuclear envelope is removed.

Tata bindingprotein TBP

38 Not suitable for samples whereDNA is removed.



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To prepare this solution weigh 5 g per 100 ml of Tris Buffer SalineTween20 (TBST, also called TBS) buffer. Mix well and filter. Failure to filtercan lead to “spotting” where tiny dark grains will contaminate the blotduring development.

Incubate for 1 hr at 4°C under agitation. Rinse for 5 sec in TBST after theincubation.

I) Incubation with primary antibodyIncubation Buffer: Dilute the antibody in TBST at the suggested dilution,if the datasheet does not have a recommended dilution try a range ofdilutions (1:100-1:3000) and optimize the dilution according to the results.Too much antibody will result in non-specific bands.It is traditional in certain laboratories to incubate the antibody in blockingbuffer, while other laboratories incubate the antibody in TBST without ablocking agent. The results are variable from antibody to antibody and youmay find it makes a difference to either use no blocking agent in theantibody buffer or the same agent as the blocking buffer.

Incubation Time: The time can vary between a few hours and overnight(rarely more than 18 hours), and is dependent on the binding affinity of theantibody for the protein and the abundance of protein. We recommend amore dilute antibody for a prolonged incubation to ensure specific binding.

Incubation Temperature: preferably cold. If incubating in blocking buffer,it is imperative to incubate at 4°C or contamination will incur and thusdestruction of the protein overnight (especially phospho groups).

Agitation of the antibody is recommended to enable adequatehomogenous covering of the membrane and prevent uneven binding.

J) Incubation with secondary antibodyIncubation Buffer: Dilute the antibody in TBST at the suggested dilution,if the datasheet does not have a recommended dilution try a range ofdilutions (1:1000- 1:20,000) and optimize the dilution according to theresults. Too much antibody will result in non-specific bands.As with the primary antibody you may choose to incubate the secondaryantibody in blocking buffer or not.

Incubation Time: 1-2 hours.

Incubation Temperature: room temperature.What secondary antibody to choose? We recommend HRP-conjugatedsecondary antibodies.ALP-conjugated secondary antibodies (alkaline phosphatase) are notrecommended as they are not sensitive enough.

Agitation of the antibody is recommended to enable adequatehomogenous covering of the membrane and prevent uneven binding.

K) Development MethodsDetection kitsFor HRP-conjugated antibodies: ECL and ECL+ (home made orcommercially available) are the traditional kits used and we recommendECL+ or more sensitive new generation kits (especially for the newgeneration detection machines such as Genegnome, etc, in those casesuse the detection kit recommended by the manufacturer of the machine).We do not recommend ECL or BCIP/NBT detection kits as they are notvery sensitive.

X-ray films: Manual film development is traditionally used and enables the scientist tocontrol the incubation time of the x-ray film in the developing agent andfixation agent.

Automated x-ray film developers are also widely used and easy to use.

Remember that an over-exposed film is not suitable foranalysis as determination of the relative amount ofprotein is not possible (overexposed films show totallyblack bands with no contrast, numerous non-specificbands, etc…).

Digital images: The new generation of film developers are light tight units with a cameradetecting the minute about of chemiluminescence emanating from the

membrane, transforming the signal into a digital image for rapid analysiswith software provided with the detection machine.A range of machines are now commercially available.

At the front of the next generation are systems which do not use HRP-conjugated antibodies (i.e chemiluminescence): for example, STORMAnalysers detect fluorescence from fluorochrome-conjugated secondaryantibodies, the Odyssey Infrared Imaging System detects infraredfluorescence.

L) Determination of the protein sizeWith SDS treatment, the proteins will migrate as a function of theirmolecular mass. A linear relationship therefore exists between the logarithm of themolecular weight of an SDS-denatured polypeptide, or native nucleic acid,and its Rf. The Rf is calculated as the ratio of the distance migrated by themolecule to that migrated by a marker dye-front. A simple way ofdetermining relative molecular weight by electrophoresis (Mr or MW) is toplot a standard curve of distance migrated vs. log 10MW for knownsamples (molecular weight markers), and read off the logMr of the sampleafter measuring the distance migrated on the same gel.

M) Western blot protocol troubleshooting tips

No signalThe primary antibody and the secondary antibody are not compatible.

Use secondary antibody that was raised against the species in which theprimary was raised (e.g primary is raised in rabbit, use anti-rabbitsecondary).

Not enough primary or secondary antibody is bound to the protein ofinterest.

Use more concentrated antibody. Incubate longer (e.g. overnight) at 4ºC.

Cross-reaction between blocking agent and primary or secondaryantibody.

Use a mild detergent such as Tween20 or switch blocking reagent (i.e.commonly used blocking reagents are milk, BSA, serum or gelatin).

The primary antibody does not recognize the protein in the speciesbeing tested.

Check the datasheet or perform a ClustalW alignment to ensure yourantibody should react with the target protein; Run the recommendedpositive control.

Insufficient antigen.

Load at least 20-30 ug protein per lane; Use protease inhibitors; Run therecommended positive control.

The protein of interest is not abundantly present in the tissue.

Use an enrichment step to maximize the signal (e.g. prepare nuclearlysates for a nuclear protein, etc.).

Poor transfer of protein to membrane.

Check the transfer with a reversible stain such as Ponceau S; check thatthe transfer was not performed the wrong way; if using PVDF membranemake sure you pre-soak the membrane in MeOH then in transfer buffer.

Excessive washing of the membrane.

Do not over wash the membrane.

Too much blocking does not allow you to visualize your protein ofinterest.

Instead of using 5% milk in the antibody buffers try removing the milk orusing 0.5%; Switch blocking reagents or block for less time.

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No signalOver-use of the primary antibody.

Use fresh antibody as the effective concentration is lowered upon each re-use.

Secondary antibody inhibited by sodium azide.

Do not use sodium azide together with HRP-conjugated antibodies.

Detection kit is old and substrate is inactive.

Use fresh substrate.

High backgroundBlocking of non-specific binding might be absent or insufficient.

Increase the blocking incubation period and consider changing blockingagent. Abcam recommends 5% non-fat dry milk, 3% BSA, or normalserum for 30 min. These can be included in the antibody buffers as well.

The primary antibody concentration may be too high.

Titrate the antibody to the optimal concentration, incubate for longer but inmore dilute antibody (a slow but targeted binding is best).

Incubation temperature may be too high.

Incubate blot at 4°C.

The secondary antibody may be binding non-specifically or reactingwith the blocking reagent.

Run a secondary control without primary antibody.

Cross-reaction between blocking agent and primary or secondary.

Add a mild detergent such as Tween20 to the incubation and washing buffer.

(phospho-specific protein) Milk contains casein which is aphosphoprotein; this is why it causes high background because thephospho-specific antibody detects the casein present in the milk.

Use BSA as a blocking reagent instead of milk.

Washing of unbound antibodies may be insufficient.

Increase the number of washes.

Your choice of membrane may give high background.

Nitrocellulose membrane is considered to give less background than PVDF.

The membrane has dried out.

Care should be taken to prevent the membrane from drying out during incubation.

Multiple bandsCell lines that have been frequently passaged gradually accumulatedifferences in their protein expression profiles.

Go back to the original non-passaged cell line and run the current andoriginal cell line samples in parallel.

The protein sample has multiple modified forms in vivo such asacetylation, methylation, myristylation, phosphorylation,glycosylation etc.

Examine the literature and use an agent to dephosphorylate, de-glycosylate, etc. the protein to bring it to the correct size.

The target in your protein sample has been digested (more likely ifthe bands are of lower molecular weight).

Make sure that you incorporate sufficient protease inhibitors in yoursample buffer.

Multiple bandsUnreported novel proteins or different splice variants that sharesimilar epitopes and could possibly be from the same protein familyare being detected.

Check the literature for other reports and also perform a BLAST search;Use the cell line or tissue reported on the datasheet.

Primary antibody concentration is too high - at high concentrationmultiple bands are often seen.

Try decreasing the antibody concentration and/or the incubation period.

Secondary antibody concentration is too high - at high concentrationsecondaries will bind non-specifically.

Try decreasing the concentration. Run a secondary antibody control(without the primary).

The antibody has not been purified.

Try to use affinity purified antibody. This will often remove non-specific bands.

The bands may be non-specific.

Where possible use blocking peptides to differentiate between specific andnon-specific bands. Only specific bands should be blocked (and thusdisappear).

The protein target may form multimers.

Try boiling in SDS-Page for 10 minutes rather than 5 minutes to disruptmultimers.

Uneven white “spots”on the blotAir bubbles were trapped against the membrane during transfer or the antibody is not evenly spread on the membrane.

Make sure you remove bubbles when preparing the gel for transfer.Incubate antibodies under agitation.

Black dots on the blotThe antibodies are binding to the blocking agent.

Filter the blocking agent.

White bands on a black blot (negative of expected blot)Too much primary and/or too much secondary antibody.

Dilute the antibodies more.

MW marker lane is blackThe antibody is reacting with the MW marker.

Add a blank lane between the MW marker and the first sample lane.

The band of interest is very low/high on the blot Separation is not efficient.

Change the gel percentage: a higher percentage for small protein, lowerpercentage for large proteins.

Smile effect of the bands1. Migration was too fast.2. Migration was too hot (changing the pH and altering the migration).

Slow down the migration or run the gel in the cold room or on ice.



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Uneven band size in lanes probed for the same protein Gel has set too quickly while casting and the acrylamide percentageis not even along the lanes.

Review the recipe of the gel and the addition of TEMED to the gels, add alittle 0.1% SDS in water to the top of the migrating gel while it sets to stopit from drying.

Uneven staining of the gel1. Contamination from bacteria2. Not enough antibody

1. Keep antibodies at 4˚C and use fresh buffers covers the gel.2. Make sure the membrane is covered with the antibody/ incubate under agitation.

N) StrippingStripping is the term used to describe the removal of primary andsecondary antibodies from the membrane. This step may remove someprotein from the membrane or damage fragile protein and a PVDFmembrane is highly recommended.

Stripping is used when more than one protein is investigated on the same blot,or the same protein with different antibodies (for example a phospho-specificantibody is probed, then the relative total amount of protein is calculated).

Three types of stripping are found. As a rule of thumb, try the gentler onefirst then proceed to harsher ones if you still get signal. Most antibodiesrequire medium or harsh stripping to remove all signal.

Gentle stripping: 10 min PBS10 min PBS20 min TBST20 min TBST

Ready for blocking stage.

Medium stripping: Make fresh stripping buffer: 15 g glycine1 g SDS10 ml Tween20Set the pH to 2.2

make up to 1 L with ultrapure water

Membrane incubation:5-10 min stripping buffer5-10 min stripping buffer10 min PBS10 min PBS5 min TBST1 min methanol rehydration (PVDF membrane only)5 min TBST


Harsh stripping: to be done under the fumehoodFor 100 ml: 20 ml SDS 10% 12.5 ml Tris HCl pH 6.8 0.5M67.5 ml ultra pure water

Add 0.8 ml ß-mercaptoethanol under the fumehood.

Make stripping buffer:Warm up the buffer at 50°C.Add the buffer to a small plastic box which has a tight lid.Add the membrane. Incubate at 50°C for up to 45 min.Dispose of the solution as required for ß-mercaptoethanol based buffers.Rinse the membrane under a running water tap for 1-2 hours.Traces of ß-mercaptoethanol will damage the antibodies. Wash extensively.1 min methanol rehydration (PVDF membrane only)5 min TBST


II. WESTERN BLOTTING OF PHOSPHO-PROTEINSPROTOCOL

Homogenize the cells or tissue of interest in lysis buffer made fresh andcontaining a cocktail of protease inhibitors (and phosphatase inhibitorswhen dealing with phosphorylated proteins).

As soon as lysis occurs, proteolysis, dephosphorylation and denaturationbegin. These events can be slowed down tremendously if samples arekept on ice or at 4°C at all times and appropriate inhibitors are addedfresh to the lysis buffer.

Use a RIPA or NP40 buffer supplemented with fresh protease andphosphatase inhibitors:

Ready to use cocktails of inhibitors from well known suppliers are oftenused but you can make your own cocktail.

Remember to add phosphatase inhibitors to cocktailsbought when investigating phosphorylation events.

Final concentrations of inhibitors: see table on page 67

1. To a sample of protein solution containing 1-100 ng of the target protein(500 ug lysate), add an equal volume of 2x SDS-PAGE sample buffer.For reduced samples, the sample buffer should be supplemented with DTTor ß-mercaptoethanol. For non-reduced samples, the DTT or ß-mercaptoethanol is not added.

2. Denature the proteins by heating the sample to 95˚C, or boiling, for 5 min.

3. Load the sample onto an SDS-polyacrylamide gel and run the gel understandard conditions.

4. Transfer the proteins to a PVDF membrane using semi-dry or wettransfer methods. Please note: for PVDF it is essential to pre-wet themembrane in methanol prior to transfer.

5. If required, the efficiency of transfer can be determined by staining themembrane briefly (10 s) in Ponceau stain. The stain can be removed bywashing in PBST or TBST. We would recommend not washing blots indistilled water as this can strip off proteins in some circumstances.

6. Block the membrane with 5% w/v BSA in TBST. Incubate for 1 h at4°C with agitation.

7. Dilute the primary antibody in TBST to the recommended dilution. Werecommend incubating in a sealed bag, hybridization tube or 50 ml Falcontubes (~2.5 ml primary antibody/blot). Incubate overnight at 4˚C withagitation.

8. Rinse the blot in TBST three to four times for 5 min each at roomtemperature.

9. Dilute the horseradish peroxidase (HRP) labeled secondary antibody atthe recommended dilution (1/5000 is usually a good working dilutionalthough this needs to be optimized for the particular application) in TBST.

10. Rinse the blot in TBST three to four times for 5 min each at roomtemperature.

11. Perform ECL Plus detection.

Top tips for WB of phosphorylated proteins:

1. Keep the proteins in their phosphorylated state! Add adequatephosphatase inhibitors and keep samples on ice at all times.2. Block the membrane in 5% w/v BSA (fractionV) NOT MILK (milkcontains casein which is a phosphoprotein; This is why it causeshigh background because the phospho-specific antibody detects thecasein present in the milk). 3. Remember the phosphorylation may need to be induced. Lowsignal or no signal may mean that the induction is not sufficient. Runthe recommended positive control with your samples.

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III. IMMUNOPRECIPITATION PROTOCOL (IP)

Immunoprecipitation is a method that enables the purification of a protein.An antibody for the protein of interest is incubated with a cell extract sothat the antibody will bind the protein in solution. The antibody/antigencomplex will then itself be pulled out of the sample using protein A/G-coupled agarose beads. This physically isolates the protein of interest fromthe rest of the sample. The sample can then be separated by SDS-PAGEfor WB analysis.

A) ReagentsLysis buffers used for IP: RIPA buffer, NP40 buffer, i.e buffers with somedetergent and with protease inhibitors (and phosphatase inhibitors iflooking at phospho-proteins).

sterile PBS pH 7.4

sterile PBS-BSA 1% (filtered)

TBST buffer

loading/sample buffer used for WB

B) Immunoprecipitation:1. On ice, in a tube add 10-500 µg cell lysate plus 0.5-5 µg of antibody.These amounts will be chosen depending on the abundance of the proteinand the affinity of the antibody for the protein, typically in a pilotexperiment where a fixed amount of protein is precipitated by increasingamounts of antibody.

2. Incubate the sample with the antibody between 1 hour to overnight(depending again on the amount of protein and affinity properties of theantibody), at 4°C, preferably under agitation.

3. Meanwhile prepare the sepharose beads. If using a monoclonalantibody choose protein G-coupled sepharose beads, if using a polyclonalantibody choose protein A-coupled sepharose beads. If the beads comeas a powder incubate 100 mg of beads in 1ml PBS 0.1M, wash for onehour so they swell up, then centrifuge, remove the supernatant anddiscard, add 1ml PBS-BSA 1% w/v, mix for one hour and rinse in PBStwice. Remove the supernatant and add 400 µl of buffer made withprotease inhibitors (can be the same as the lysis buffer). The slurry is nowready for use. It can be stored at 4°C for a few days; for longer periodskeep the beads in PBS with 0.02% azide (rinse extensively the beads onthe day of use and make up in lysis buffer). You can also buy pre-swollenbeads as slurry ready for use.

IgM antibody: Do not use protein-A or protein-Gconjugated beads, use Goat anti Mouse IgM (orpolyvalent Ig, or anti-heavy chain) beads.

4. Mix the slurry well and add 70-100 ul of the beads to each sample. Alwayskeep samples on ice. Beads will tend to stick to the sides of the tip so try tominimize the movement in the pipette and use a tip cut 5 mm from the top.

5. Incubate the lysate-beads mixture at 4°C under rotary agitation for 4hours (the optimal incubation time can be determined in a preliminaryexperiment).

6. When the incubation time is over, centrifuge the tubes, remove thesupernatant and wash the beads in lysis buffer three times (each timecentrifuging at 4°C and removing the supernatant).

7. Finally, remove the last supernatant and add 25-50 µl of 2x loadingbuffer. Boil at 95-100°C for 5 minutes to denature the protein and separateit from the protein-A/G beads, then centrifuge and keep the supernatantwhere the protein is now. You can then freeze the samples or run them ona SDS-PAGE. See WB protocol for further details (page 66).

Choosing the protein beads: Summary

IV. NUCLEAR FRACTIONATION PROTOCOL

A) ReagentsBuffer A – 10 mM HEPES, 1.5 mM MgCl2, 10 mM KCl, 0.5 mM DTT,0.05% NP40 (or 0.05% Igepal or Tergitol) pH 7.9

To prepare 250 ml stock of buffer A – HEPES: 1M = 238.3 g/L, therefore 10 mM = 0.59 g/250mlMgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250mlKCl: 1M = 74.5 g/L, therefore 10 mM = 0.187 g/250mlDTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250mlNP40 = 0.05%

Buffer B – 5 mM HEPES, 1.5 mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT,26% glycerol (v/v), pH 7.9

To prepare 250 ml stock of buffer B –HEPES: 1M = 238.3 g/L, therefore 5mM = 0.295 g/250mlMgCl2: 1M = 203.3 g/L, therefore 1.5mM = 0.076 g/250mlEDTA: 1M = 372.2 g/L, therefore 0.2mM = 0.0186 g/250mlDTT: 1M = 154.2 g/L, therefore 0.5mM = 0.019 g/250ml26% Glycerol (v/v) = 65ml

4.6 M NaCl – 87.66 g/326ml

Species Immunoglobulin Isotype Protein A Protein G

Human IgG1 333 333

IgG2 333 333

IgG3 - 333

IgG4 333 333

IgM Use anti Human IgM

IgE - 3

IgA - 3

Mouse IgG1 3 333

IgG2a 333 333

IgG2b 33 33

IgG3 3 3

IgM Use anti Mouse IgM

Rat IgG1 - 3

IgG2a - 333

IgG2b - 33

IgG2c 3 33

Chicken All isotypes - 33

Cow All isotypes 33 333

Goat All isotypes - 33

Guinea Pig All isotypes 333 33

Hamster All isotypes 3 33

Horse All isotypes 33 333

Pig All isotypes 33 33

Rabbit All isotypes 333 33

Sheep All isotypes - 33



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B) Method1. Prepare 1 ml of buffer A with added cocktail of usual inhibitors fromfrozen stock and store on ice.

2. Add 500 µl of buffer a per large petri dish on ice and scrape thoroughly,leave on ice for 10 min.

3. Centrifuge at 4˚C at 3000 rpm for 10 min.

4. Remove supernatant and keep it (this will contain everything exceptlarge plasma membrane pieces, DNA, nucleoli), extract out 10 µl forBradford assay.

5. On ice resuspend pellet in 374 µl of buffer B and add 26 µl of 4.6 MNaCl to give 300mM NaCl (high salt helps lyse membranes and forcesDNA into solution).

6. Homogenize with 20 full strokes in Dounce or glass homogenizer on ice.

7. Leave on ice for 30 min.

8. Centrifuge at 24,000 g for 20 min at 4˚C.

9. Aliquot supernatant, remove 10 µl for Bradford assay and store at -70˚C.

V. MITOCHONDRIAL PURIFICATION PROTOCOL

This protocol is for use with cells in suspension.

1. Collect cells by centrifugation at approximately 370 g for 10 minutes.Decant supernatant and re-suspend cells in 10 packed cell volumes ofNKM buffer (1 mM TrisHCl, pH 7.4, 0.13 M NaCl, 5 mM KCl, 7.5 mM MgCl2).

2. Pellet cells and decant supernatant, repeat this washing step 2 times.Resuspend cells in 6 packed cell volumes of homogenization buffer (10mM Tris-HCl, pH 6.7, 10 mM KCl, 0.15 mM MgCl2, 1 mM PMSF, and 1 mMDTT, always add PMSF and DTT immediately before use).

3. Transfer cells to a glass homogenizer and incubate for 10 minutes onice. Using a tight pestle, homogenize the cells. Check under themicroscope for cell breakage, the optimum is around 60%. This mayrequire 30 strokes or so of the pestle.

4. Pour homogenate into a conical centrifuge tube containing 1 packed cellvolume of 2 M sucrose solution and mixed gently. Pellet unbroken cells,nuclei, and large debris at 1200 g for 5 minutes and transfer thesupernatant to another tube. This treatment is repeated twice, transferringthe supernatant to a new tube each time, discarding the pellet.

5. Pellet the mitochondria by centrifuging at 7000 g for 10 minutes.Resuspend the mitochondrial pellet in 3 packed cell volumes ofmitochondrial suspension buffer (10 mM TrisHCl. pH 6.7, 0.15 mM MgCl2,0.25 mM sucrose, 1 mM PMSF, 1 mM DTT). Spin at 9500 g for 5 minutesto re-pellet the mitochondria.

6. At this point, you can add 1X protein gel loading buffer and run on a gelif a whole mitochondrial protein extract is needed, further purify themitochondria on a sucrose gradient if you really need very puremitochondria, or purify a soluble (S-100) fraction of the mitochondria. Thislast protocol is given below.

VI. SOLUBLE (S-100) MITOCHONDRIALFRACTIONATION PROTOCOL

1. Resuspend mitochondria in one-third the packed cell volumemitochondrial lysis buffer (25 mM HEPES-KOH pH 7.6, 5 mM MgCl2, 0.5mM EDTA, 10% glycerol, 1 mM DTT, 1 mM PMSF). Put the suspensioninto a glass homogenizer and homogenize with 10 strokes using a tightpestle. Add Tween20 and KCl to final concentrations of 0.5% and 0.5 M,respectively.

2.Incubate the mixture on ice 5 minutes. The homogenization is repeatedfor a total of 10 times.

3. Spin the final mitochondrial lysate at 100,000 g in an ultracentrifugeusing a TY65 Beckman rotor at 4˚C, for 60 minutes.

4. Carefully collect the clear supernatant, avoiding the fluffy layer over thepellet, to yield the final S-100 fraction.

5. Freeze in aliquots, in liquid nitrogen, and store at -80˚C.

This is a slightly modified protocol taken from Methods in Enzymology, Vol264, by Vicente Micol, Patricio Fernandez-Silva, and Giuseppe Attardi.

VII. DETERMINING IF THE ANTIBODY BINDS ONLYPHOSPHORYLATED PROTEINS (WB OR IHC) PROTOCOL

1. Tissue sections or PVDF membranes (after protein transfer andrehydratation of the membrane in 100% methanol followed by water)should be incubated overnight at 37°C in 1% calf intestinal alkalinephosphatase (CIP, New England Biolabs, Beverly, MA, USA) diluted in NEbuffer 1 (in mM: NaCl, 10; Tris, 5, pH 7.9; MgCl2, 1; dithiothreitol, 0.1).

2. Sister sections or membranes should be incubated in the NE buffer only.Rinse three times in PBST (IHC) or TBST (WB) the sections or membranes,respectively.

3. In PBST (IHC) or TBST (WB), membranes in 5% milk.

4. Incubate sections and membranes overnight in primary antibody.

5. Secondary antibodies concentrations and revelation protocol should beperformed as usual.

Ref: European Journal of Neuroscience, (2005) 21 (7) pp. 1785–97, Sophie Pezet, Achillefs Spyropoulos, Robert J. Williams and Stephen B.McMahon.

VIII. DOT BLOT PROTOCOL

A technique for detecting, analyzing, and identifying proteins, similar to thewestern blot technique but differing in that protein samples are notseparated electrophoretically but are spotted through circular templatesdirectly onto the membrane or paper substrate.

Concentration of proteins in crude preparations (such as culturesupernatant) can be estimated semi-quantitatively by using “Dot Blot”method if you have both purified protein and specific antibody against it.

A) ReagentsTBS: 20 mM Tris-HCl, 150 mM NaCl, pH 7.5TBS-T: 0.05% Tween20 in TBSBSA/TBS-T: 0.1% BSA in TBS-TNitrocellulose membrane (BIO-RAD, Trans-Blot, etc.)

B) Procedure1. Have nitrocellulose membrane ready, draw grid by pencil to indicate theregion you are going to blot (see below).

2. Using narrow-mouth pipet tip, spot 2 µl of samples onto thenitrocellulose membrane at the center of the grid. Minimize the area thatthe solution penetrates (usually 3-4 mm diam.) by applying it slowly.

3. Let the membrane dry.

4. Block non-specific sites by soaking in 5% BSA in TBS-T (0.5-1 hr, RT).Use 10cm Petri Dish for reaction chamber.

5. Incubate with primary antibody (0.1-10 µg/ml for purified antibody,1:1000 to 1:100000 dilution for antisera, 1:100 to 1:10000 for hybridomasupernatant) dissolved in BSA/TBS-T for 30 min at RT.

6. Wash three times with TBS-T (3 x 5 min).

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7. Incubate with secondary antibody conjugated with HRP (for optimumdilution, follow the manufacturer’s recommendation) for 30 min at RT.

8. Wash three times with TBS-T (15 min x 1, 5 min x 2), then once withTBS (5 min).

9. Incubate with ECL reagent for 1 min, cover with Saran-wrap (removeexcessive solution from the surface), and expose X-ray film in the darkroom. Try several different lengths of exposure.

10. Compare the signal from your unknown sample to that of standard andestimate the concentration.

IX. BLOCKING WITH IMMUNIZING PEPTIDE (BL)PROTOCOL

It is not uncommon to see more than one band in Western blotting whenprobing with a given antibody or to see more diffuse staining inimmunolocalization studies. The question arises which band or staining isspecific.

The antibody specificity is generally studied by competing with excessantigen (peptide or protein) or immuno-neutralization with the antigens.

In principle, a small volume of antibody (e.g. 1-5 µl) is first reacted withexcess peptide (5-50 fold over the antibody; e.g. 1 µg antibody reactedwith 5-50 µg peptide; exact amounts determined by titration) to neutralizeit. The neutralized antibody can no longer bind to another antigen. So theband(s)/staining that is competed by the antigen/peptide is specific. If morethan one band disappears by peptide/antigen competition then thosebands have the antigenic determinants and could be considered eitherfragments of the large antigen or multimer.

1. Determine the amount of antibody that is needed for 1 strip (e.g., 2 ml).For example, an antibody has given desired bands at 1:1000 dilution. Soyou will need 1 µl/ml antibody (2 µl antibody for 2 ml antibody solution). Ifan antibody was used at 1:5000 dilution then you would only need 0.2µl/ml (use 2 µl of 1:10 dilution for better accuracy).

2. Take 2 µl antibody (or as needed) in 100 µl saline/PBS. Make 2 tubes.Add antigen/peptide solution (10-50 µg peptide or antigen added in 10-100µl). Add same volume of saline/PBS (no peptide/antigen) to the other tubelabeled as “no peptide”. Mix gently.

3. Incubate both tubes at 37˚C for 1-2 hrs or 2-24 hrs at 4˚C.

4. Centrifuge the tubes for 15 min at 4˚C in a microfuge (10-15000 rpm) topellet any immune complexes. Carefully remove the supernatant. If novisible pellet is seen then just leave ~ 5-10 µl at the bottom to avoiddisturbing invisible immune complexes. If you do not centrifuge thesolution, it may give high background.

5. After centrifugation, make up the volume of supernatant to 2 ml (or whatis necessary depending upon the initial antibody taken) with buffer (PBS-Tween with or without BSA or milk or any buffer that was used initially).Use both antibodies (with and without antigen/peptide) for Western blottingor immunolocalization.

6. Observe the bands/staining that disappears.

X. IHC-PARAFFIN PROTOCOLS (IHC-P)

A) Antigen retrievalFormalin fixed tissue requires an antigen retrieval step beforeimmunohistochemical staining can proceed. This is due to the formation ofmethylene bridges during fixation, which cross link proteins and thereforemask antigenic sites. The two methods of antigen retrieval are enzymaticand heat mediated. Both serve to break the methylene bridges and soexpose the antigenic sites in order to allow the antibodies to bind. Someantigens prefer enzymatic to heat mediated antigen retrieval and viceversa. Enzymatic tends to be a much gentler process than heatedmediated, so is best suited to more friable tissues. However, enzymatictends to take longer and is more technically demanding.If no antigen retrieval step is stated on the antibody data sheet, start off bytrying the heat mediated pressure cooker method.

i) Enzymatic Antigen RetrievalTissue sections are best mounted on APES (amino-propyl-tri-ethoxy-silane) coated slides. Slides should be placed in a rack for this procedure.

Materials• Water baths • 2 troughs • pH meter • Magnetic stirrer • Thermometer

Reagents• Alpha-Chymotrypsin (type II from Bovine pancreas) 0.1 g • Calcium Chloride 0.1 g • Ultra-pure water 100.0 ml • 0.5% sodium hydroxide solution • 0.5% hydrochloric acid solution • Xylene • Industrial methylated spirits (IMS) or methanol

Method1. Set water bath to 37°C. Add the required amount of ultra pure water intoeach trough then place the troughs into the water bath (See note i). Allowthe ultra pure water to warm to 37°C.

2. De-wax and re-hydrate paraffin sections by placing them in 3 changesof xylene for 3 minutes each, followed by 3 changes of IMS or methanolfor 3 minutes each, followed by cold running tap water for 3 minutes. At notime from this point onwards should the slides be allowed to dry out! Placeslides in one trough of ultra pure water at 37°C to warm (See note ii).

3. Remove the other trough and into this dissolve the calcium chloride andchymotrypsin using a magnetic stirrer (See note iii). Once dissolved, pH to7.8 using the 0.5% sodium hydroxide and hydrochloric acid solutions.Return the trough to the water bath and allow this enzyme solution to re-heat to 37°C (See note iv).

4. Transfer the warmed slides into the enzyme solution for a suggested 20minutes (See note v) then remove the slides and place them into coldrunning tap water for 3 minutes (See note vi).

5. Continue with immunohistochemical staining protocol.

Notesi. Use a sufficient volume of ultra pure water in order to cover the slides.Adjust the reagent quantities appropriately (the recipe above in the“Reagents” section is for 100 ml).

ii. Placing cold slides into the enzyme solution will lower the temperature ofthe solution, therefore reducing enzyme activity leading to the antigensbeing under-retrieved. Once the slides have been de-waxed and re-hydrated, drying out will cause non-specific antibody binding and thereforehigh background staining.

iii. Chymotrypsin can be very allergenic. Use a face mask and extractioncabinet for weighing out.

iv. Prepare the chymotrypsin solution as quickly as possible to avoidimpairing the activity of the enzyme. Allow this solution to return to 37°Cbefore introducing the slides.

v. Twenty minutes is only suggested as a starting point incubation time.Less than 20 minutes may leave the antigens under retrieved, leading toweak staining. More than 20 minutes may leave them over retrieved,leading to non-specific background staining and also increasing thechances of sections dissociating from the slides. A control experiment isrecommended beforehand, where slides of the same tissue section areincubated in the enzyme solution for 10, 15, 20, 25, and 30 minutes beforebeing immunohistochemically stained to evaluate optimum antigenretrieval time for the particular antibody being used.

vi. Tap water stops the enzymatic process by washing action.

ii) Pressure Cooker MethodTissue sections are best mounted on APES (amino-propyl-tri-ethoxy-silane) coated slides. Slides should be placed in a metal rack for thisprocedure.



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Materials• Domestic stainless steel pressure cooker • Hot plate • pH meter • Magnetic stirrer • 2 L beaker or conical flask

Reagents• Tri-Sodium Citrate 5.88 g • 0.2 M Hydrochloric acid solution 44 ml • Ultra-pure water 1956 ml • 0.5% sodium hydroxide solution • 0.5% hydrochloric acid solution • Xylene • Industrial methylated spirits (IMS) or methanol

Method1. Add the tri-sodium citrate, hydrochloric acid and ultra-pure watertogether in a 2 L beaker/conical flask. Use a magnetic stirrer to ensure thatall reagents are properly dissolved.

2. Adjust to pH 6.0 with 0.5% sodium hydroxide and hydrochloric acidsolutions. Add this solution to the pressure cooker. Place the pressurecooker on the hotplate and turn it on full power. Do not secure the lid ofthe pressure cooker at this point, simply rest it on top.

3. While waiting for the pressure cooker to come to a boil, de-wax and re-hydrate the paraffin sections by placing them in 3 changes of xylene for 3minutes each, followed by 3 changes of IMS or methanol for 3 minuteseach, followed by cold running tap water. Keep them in the tap water untilthe pressure cooker comes to a boil. At no time from this point onwardsshould the slides be allowed to dry out! (See note i.)

4. Once boiling, transfer the slides from the tap water to the pressurecooker. USE CARE WITH HOT SOLUTION - USE FORCEPS! Secure thepressure cooker lid as in the manufacturer’s instructions.

5. As soon as the cooker has reached full pressure (see the manufacturersinstructions), time 3 minutes. (See note ii).

6. When 3 minutes has elapsed, turn off the hotplate and place thepressure cooker in an empty sink. Activate the pressure release valve (seethe manufacturer’s instructions) and run cold water over the cooker. Oncede-pressurized, open the lid and run cold water into the cooker for 10minutes. CARE WITH HOT SOLUTION! (See note iii).


Notesi. Once the slides have been de-waxed and re-hydrated, drying out willcause non-specific antibody binding and therefore high backgroundstaining.

ii. 3 minutes is only suggested as a starting point antigen retrieval time.Less than 3 minutes may leave the antigens under retrieved, leading toweak staining. More than 3 minutes may leave them over retrieved,leading to non-specific background staining and also increasing thechances of sections dissociating from the slides. A control experiment isrecommended beforehand, where slides of the same tissue section areretrieved for 1, 2, 3, 4 and 5 minutes before being immunohistochemicallystained to evaluate optimum antigen retrieval time for the particularantibody being used.

iii. This is to make the slides cool enough to handle and to allow theantigenic site to re-form after being exposed to such high temperature.

iii) Microwave MethodThe use of a domestic microwave is inadvisable. Hot and cold spots arecommon, leading to uneven antigen retrieval. Antigen retrieval times areusually longer, due to the absence of a pressurized environment, nearlyalways leading to section dissociation.A scientific microwave is much more appropriate. Most brands have on-board pressurized vessels and can keep the temperature at a constant98°C to avoid section dissociation. The only drawback is the expense ofpurchasing one! Tissue sections are best mounted on APES (amino-propyl-tri-ethoxy-silane) coated slides. Slides should be placed in a plastic rack for thisprocedure.

Materials• Domestic (850W) or scientific microwave • pH meter • Magnetic stirrer • 1 L beaker or conical flask • Microwaveable vessel, either in-built or to hold approximately 400-500 ml

Reagents• Tri-Sodium Citrate 2.94 g • 0.2 M Hydrochloric acid solution 22 ml • Ultra-pure water 978 ml • 0.5% sodium hydroxide solution • 0.5% hydrochloric acid solution • Xylene • Industrial methylated spirits (IMS) or methanol

Method1. De-wax and re-hydrate the paraffin sections by placing them in 3changes of xylene for 3 minutes each, followed by 3 changes of IMS ormethanol for 3 minutes each, followed by cold running tap water. Keepthem in the tap water until the microwave antigen retrieval solution hasbeen prepared. At no time from this point onwards should the slides beallowed to dry out! (See note i)

2. Add the tri-sodium citrate, hydrochloric acid and ultra-pure watertogether in a 1 L beaker/conical flask. Use a magnetic stirrer to ensure thatall reagents are properly dissolved. Adjust to pH 6.0 with 0.5% sodiumhydroxide and hydrochloric acid solutions. Add this solution to themicrowaveable vessel (See note ii).

3. Remove the slides from the tap water and place them in themicrowaveable vessel. Place the vessel inside the microwave. If domestic,set to full power and wait until the solution comes to the boil. Boil for 15minutes from this point. If scientific, program so that antigens are retrievedfor 15 minutes once the temperature has reached 98°C. (See note iii).

4. When 15 minutes has elapsed, remove the vessel and run cold tapwater into it for 10 minutes. USE CARE WITH HOT SOLUTION! (See note iv).


Notesi. Once the slides have been de-waxed and re-hydrated, drying out willcause non-specific antibody binding and therefore high background staining.

ii. Use a sufficient volume of antigen retrieval solution in order to cover theslides. This should be by at least a few cm if using a non-sealed vessel toallow for evaporation during the boil.

iii. 15 minutes is only a suggested antigen retrieval time. Less than 15minutes may leave the antigens under retrieved, leading to weak staining.More than 15 minutes may leave them over retrieved, leading to non-specific background staining and also increasing the chances of sectionsdissociating from the slides. A control experiment is recommendedbeforehand, where slides of the same tissue section are retrieved for 5,10, 15, 20, 25 and 30 minutes before being immunohistochemicallystained to evaluate optimum antigen retrieval time for the particularantibody being used.

iv. This is to make the slides cool enough to handle and to allow theantigenic site to re-form after being exposed to such high temperature.

B) ABC Immunohistochemical Staining -Chromogenic detectionGeneral GuidelinesCarry out all incubations in a humidified chamber so that the sections donot dry out. At no stage during the protocol should drying of the sectionsbe allowed to occur. Drying out will lead to non-specific binding andultimately high background staining. A shallow, plastic “sandwich type” boxwith a sealed lid and wet tissue paper in the bottom will be adequate, justas long as the slides can lay flat so that the reagents don’t drain off!

A better solution is to cut a plastic serological pipette into lengths to fit yourincubation chamber. Glue them in pairs to the bottom of the chamber, withthe 2 individual pipette tubes of each pair being placed about 4.0 cm apart.This provides a level and raised surface for the slides to rest on away fromthe wet tissue paper.

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Carry out all TBS rinses between reagents in an appropriate glass trough.

Dilutions of the primary antibody, secondary antibody and the ABCcomplex should be determined by the individual product’s data sheet orfrom past experience. Adjust dilutions appropriately from the resultsobtained.

Adhere strictly to all incubation times in the protocol

Note: The main body of this protocol focuses on an ABC-Enzymaticmethod (3 step). However, this protocol can be easily adapted for an ABC-Fluorescent method and also a 2 step or 1 step enzymatic or fluorescentmethod. See the “Protocol Adaptations” section for details.

ProtocolPlease refer to Notes section for the theory and the Reagents section forthe recipes:

Perform antigen retrieval before commencing with thefollowing protocols!

DAY 1:1. Rinse 2 x 5min TBS 2. 1.6% H2O2 in TBS for 30 min (HRP only) (See note i) 3. Rinse 2 x 5min TBS 0.025% Triton (See note ii) 4. Block in 10% NS with 1% BSA in TBS for 2 hours at room

temperature (See note iii) 5. Drain slides for a few seconds (do NOT rinse) and wipe round

sections (See note iv) 6. Apply primary antibody made up in TBS with 1% BSA. (See note v) 7. Incubate overnight at 4˚C, preferably on an orbital shaker

(GENTLY!) (See note vi)

DAY 2:8. Rinse 2 x 5min TBS 0.025% Triton 9. Apply secondary biotinylated antibody made up in TBS with 1% BSA

for 2 hours at room temperature (AT THIS POINT MAKE UP THE ABC COMPLEX IN TBS) (See note vii and viii)

10. Rinse 2 x 5min TBS 11. Apply ABC complex in TBS for 30 min at room temperature (See note ix) 12. Rinse 2 x 5min TBS 13. Develop with chromogen for 10 min at room temperature (See note x)14. Rinse in running tap water for 5 min. 15. Counterstain (If required) 16. Dehydrate, clear and mount (See note xi and xii)

Notesi. H2O2 suppresses endogenous peroxidase activity and therefore reducesbackground staining. Using a low concentration for 10 minutes adequatelyblocks endogenous peroxidase activity without having a detrimental effecton tissue epitopes.

If using AP, then omit this step and step 1). See note x for further details.

ii. The use of 0.025% Triton in the TBS helps to reduce surface tension,allowing reagents to cover the whole tissue section with ease. It is alsobelieved to dissolve Fc receptors, therefore reducing non-specific binding.

Abcam recommends TBS to give a cleaner background than PBS iii. The secondary antibody may cross react with endogenousimmunoglobulins in the tissue. This is avoided by pre-treating the tissuewith normal serum from the species in which the secondary was raised.

For example, when detecting CD4 on mouse tissue and the secondary israised in a goat:

Firstly, block in normal goat serum. This will bind to any mouseimmunoglobulins that show cross reactivity with goat immunoglobulins.

On addition of the primary antibody, for example a Rabbit IgG anti-MouseCD4, this antibody will bind to its target epitope on the CD4 molecule.

On addition of the secondary antibody, for example a biotinylated Goatanti-Rabbit IgG, this antibody will now only bind to the primary Rabbit IgGanti-Mouse CD4 antibody.

If the secondary antibody did have an affinity for any of the endogenousimmunoglobulins on the mouse tissue, it can no longer bind to them asthey have already been bound to immunoglobulins in the goat serum.Immunoglobulins from the same species will not interact with each otherand therefore the secondary can only bind to the primary antibody.

The use of normal serum before the application of the primary alsoeliminates Fc receptor binding of both the primary and secondary antibody.

The BSA serves a similar purpose as the normal serum, but this works toreduce non-specific binding caused by hydrophobic interactions.

Cases can occur where the primary antibody recognizes another epitopeshowing sufficient homology with its actual target, for example a non-related tissue component. This type of background is greatly reduced bythe careful screening of monoclonal antibodies and the absorption ofpolyclonal antibodies. General non-specific binding of Abcam primaryantibodies is now rare for similar reasons.

Monoclonal antibodies tend to show less cross-reactivity with closelyhomologous epitopes than polyclonals, due to each monoclonal antibodymolecule recognizing exactly the same epitope.

iv. Simply remove the excess serum so not to further dilute the primaryantibody.

v. The primary antibody will target the epitope it is raised against. Forexample, an amino acid sequence unique to the CD4 molecule.

Make sure that the primary antibody is raised in a different species to thetissue being stained. If, for example, you had mouse tissue and yourprimary antibody was raised in a mouse, the application of an anti-mouseIgG secondary antibody would bind to all the endogenous IgG in themouse tissue. High non-specific background would occur. As discussedearlier, pre-incubation of the tissue with normal mouse serum would haveno effect due to immunoglobulins from the same species not interactingwith each other

vi. Overnight incubation allows antibodies of lower titer or affinity to beused by simply allowing more time for the antibodies to bind. Also,whatever the antibody’s titer or affinity for its target, once the tissue hasreached saturation point no more binding can take place. Overnightincubation assures that this occurs.

The lower incubation temperature and gentle agitation from an orbitalshaker is believed to help reduce background staining by increasingreaction times.

vii. The secondary antibody recognizes the immunoglobulin species andsubtype of the primary antibody.

In this example, a biotinylated Goat anti-Rabbit IgG antibody is being usedto bind to the primary Rabbit IgG anti-Mouse CD4 antibody.

The secondary antibody is biotinylated, meaning that it has beenconjugated with biotin.

When applied, the ABC complex will bind to this secondary biotinylatedantibody.

viii. After the addition of the secondary antibody, make up the ABCcomplex and leave to stand for a minimum of 30 minutes. This is thelength of time that the complex takes to form.

ix. The ABC complex consists of biotin-HRP/AP conjugate bound to avidin.

Avidin is a protein found in chicken egg white and has similar properties tostreptavidin, a protein found in Streptomyces avidinii. Both Avidin andstreptavidin have a high affinity for biotin, a co-factor in enzymes involvedin carboxylation reactions.

Both avidin and streptavidin can be used in an IHC application, butstreptavidin tends to be favored since it shows greater sensitivity.Streptavidin also produces less non-specific background staining, due to itbeing non-glycosylated (unlike avidin), so shows no interaction with lectinsor other carbohydrate binding proteins.



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The complex is formed through avidin having 4 binding sites for biotin. Dueto steric hindrance, only 3 or fewer biotin molecules can actually bind. TheABC complex binds to the biotinylated secondary antibody, which is boundto the primary antibody, which is in turn bound to the target on the tissuesection.

Due to the lack of BSA, the surface tension of the ABC solution is greaterthan the other reagents and will need spreading out over the tissue. A 200µl pipette tip is good for this.

x. Develop the colored product of the enzyme with the appropriatechromogen. The choice depends on which enzyme label you are using,the colored end product you prefer and whether you are using aqueous ororganic mounting media (See note xi) for further details):

xi. Don’t forget that DAB is a suspected carcinogen. Wear the appropriateprotective clothing. Deactivate it with chloros in a sealed containerovernight (it produces noxious fumes when chloros is added) and disposeof it according to laboratory guidelines.

If using AP, add 0.24 mg/ml Levamisole (Sigma L9756) to the chromogensolution. Levamisole suppresses endogenous phosphatase activity andtherefore reduces background staining.

xii. If using AEC, Fast Red, INT or any other aqueous chromogen thendon’t forget that they are alcohol soluble. Use a suitable aqueousmounting media. Don’t dehydrate and clear!

xiii. Dehydrate and clear DAB, New Fuchsin, Vega Red, NBT, TNBT or anyother organic chromogen developed sections by sending them through 3changes of methanol (or IMS) for 3 minutes each followed by 3 changes ofxylene for 3 minutes each. Mount sections in a suitable organic mountingmedia. Sections mounted in organic mounting media have a betterrefractive index than those mounted in aqueous mounting media. Thismeans that the image seen down the microscope will be sharper andclearer if organic mounting media is used.

ConclusionYou should now see a colored product localized at the site of antibodybinding.

This corresponds to the location of your target.

C) ABC Immunohistochemical staining - fluorescentdetection

Perform antigen retrieval before commencing with thefollowing protocols!

DAY 1:1. Rinse 3 x 5min PBS 2. Rinse 2 x 5min PBS 0.025% Triton3. Apply primary antibody - enzyme conjugate made up in PBS with

1% BSA4. Incubate overnight at 4˚C, preferably on an orbital shaker

DAY 2:6. Rinse 3 x 5min PBS 7. Incubate with secondary anitbody which is fluorescently labelled8. Rinse 3 x 5min PBS9. Mount

D) IHC troubleshooting tips

No stainingThe primary antibody and the secondary antibody are not compatible.

Use secondary antibody that was raised against the species in which theprimary was raised (e.g primary is raised in rabbit, use anti-rabbit secondary).

Not enough primary antibody is bound to the protein of interest.

Use less dilute antibody, Incubate longer (e.g. overnight) at 4˚C.

The antibody may not be suitable for IHC procedures which reveal theprotein in its native (3D form).

Test the antibody in a native (non-denatured) WB to make sure it is notdamaged.

The primary/secondary antibody/amplification kit may have lost itsactivity due to improper storage, improper dilution or extensivefreezing/thawing.

Run positive controls to ensure that the primary/secondary antibody isworking properly.

The protein is not present in the tissue of interest.

Run a positive control recommended by the supplier of the antibody.

The protein of interest is not abundantly present in the tissue.

Use an amplification step to maximize the signal.

The secondary antibody was not stored in the dark.

Always prevent the secondary antibody from exposure to light.

Deparafinization may be insufficient.

Deparaffinize sections longer, change the xylene.

Fixation procedures (using formalin and paraformaldehyde fixatives)may be modifying the epitope the antibody recognizes.

Use antigen retrieval methods to unmask the epitope, fix for less time.

The protein is located in the nucleus and the antibody (Nuclear protein)cannot penetrate the nucleus.

Add a permeabilizing agent to the blocking buffer and antibody dilution buffer.

The PBS buffer is contaminated with bacteria that damage thephosphate groups on the protein of interest.

Add 0.01% azide in the PBS antibody storage buffer or use fresh sterilePBS.

High background - Possible reasons and solutionsBlocking of non specific binding might be absent or insufficient.

Increase the blocking incubation period and consider changing blockingagent. Abcam recommends 10% normal serum 1hr for sections or 1-5%BSA for 30 min for cells in culture.

The primary antibody concentration may be too high.

Titrate the antibody to the optimal concentration, incubate for longer but inmore dilute antibody (a slow but targeted binding is best).

Incubation temperature may be too high.

Incubate sections or cells at 4°C.

The secondary antibody may be binding non-specifically (damaged).

Run a secondary control without primary antibody.

HRP AP

AEC (red) Fast Red (pink)

DAB (brown) INT (yellow/brown)

NBT (purple/brown)

New Fuchsin (red)

TNBT (purple)

Vega Red (pink)

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High backgroundTissue not washed enough, fixative still present.

Wash extensively in PBS between all steps.

Endogenous peroxidases are active.

Use enzyme inhibitors i.e. Levamisol (2 mM) for alkaline phosphatase orH2O2 (0.3% v/v) for peroxidase. (See IHC protocol).

Fixation procedures (using formalin and paraformaldehyde fixatives)are too strong and modified the epitope the antibody recognizes.

Change antigen retrieval method, decrease the incubation time with theantigen unmasking solution.

Too much amplification (Amplification technique).

Reduce amplification incubation time and dilute the amplification kit.

Too much substrate was applied (enzymatic detection).

Reduce substrate incubation time.

The chromogen reacts with the PBS present in the cells/tissue(enzymatic detection).

Use Tris buffer to wash sections prior to incubating with the substrate, thenwash sections/cells in Tris buffer.

Pemeabilization has damaged the membrane and removed themembrane protein (membrane protein).

Remove permeabilizing agent from your buffers.

Non-specific stainingPrimary/secondary antibody concentration may be too high.

Try decreasing the antibody concentration and/or the incubation period.Compare signal intensity against cells that do not express the target.

Endogenous peroxidases are active.

Use enzyme inhibitors i.e. Levamisol (2 mM) for alkaline phosphatase orH2O2 (0.3% v/v) for peroxidase. (See IHC protocol).

The primary antibody is raised against the same species as thetissue stained (e.g Mouse primary antibody tested on mouse tissue).When the secondary antibody is applied it binds to all the tissue as itis raised against that species.

Use a primary antibody raised against a different species than your tissue.

The sections/cells have dried out.

Keep sections/cells at high humidity and do not let them dry out.

XI. IMMUNOHISTOCHEMISTRY (IHC-FR) - FROZENSECTIONS PROTOCOL

Frozen sections: Once mounted on APES coated slides, frozen sectionsthey are best kept at -80˚C until needed.

1. When required, leave to warm at room temperature for 5 min.

2. Pre-cool the fixative (acetone, methanol or ethanol) (Abcamrecommends starting with acetone) at -20˚C for 30 min.

3. Fix with the pre-cooled fixative for 5 -10 min, at room temperature

4. Rinse 3-4 X in PBS.

5. Continue with the immunohistochemical staining protocol. The absenceof formalin eliminates the need for an antigen retrieval step. However,if frozen tissue or cytological specimens have been fixed in formalin,antigen retrieval can be attempted although the friable nature of thespecimens may compromise the success.

See section X, A) and B) for detailed protocols for chromogenic andfluorescent detection.

XII. IMMUNOCYTOCHEMISTRY (ICC) PROTOCOL

i Coat coverslips with polyethylineimine or poly-L-lysine for 1 hr at room temperature.

ii Rinse coverslips well with sterile H2O (3 times 5 min each). iii Allow coverslips to dry completely and sterilize them under UV light for

at least 4 hrs. iv Grow cells on glass coverslips or prepare cytospin or smear preparation. v Rinse briefly in phosphate-buffered saline (PBS).

Fixation:1. Fix the samples either in ice-cold methanol, acetone (1-10 min) or in 3-4% paraformaldehyde in PBS pH 7.4 for 15 min at room temperature.

2. Wash the samples twice with ice cold PBS.

Permeabilization:If the target protein is expressed intracellularly, it is very important topermeabilize the cells. Note: acetone fixed samples do not requirepermeabilization.

3. Incubate the samples for 10 min with PBS containing 0.25% Triton X-100 (or 100 µM digitonin or 0.5% saponin). Triton X-100 is the mostpopular detergent for improving the penetration of the antibody. However, itis not appropriate for the use of membrane-associated antigens since itdestroys membranes.

4. Wash cells in PBS three times for 5 min.

Blocking and Incubation:5. Incubate cells with 1% BSA in PBST for 30 min to block unspecificbinding of the antibodies (alternative blocking solutions are 1% gelatin or10% serum from the species that the secondary antibody was raised in).

6. Incubate cells in the diluted antibody in 1% BSA in PBST in a humidifiedchamber for 1 hr at room temperature or overnight at 4˚C.

7. Decant the solution and wash the cells three times in PBS, 5 min eachwash.

8. Incubate cells with the secondary antibody in 1% BSA for 1 hr at roomtemperature in dark.

9. Decant the secondary antibody solution and wash three times with PBSfor 5 min each in dark.

Counter staining:10. Incubate cells on 0.1-1 µg/ml Hoechst or DAPI (DNA stain) for 1 min.

11. Rinse with PBS.



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Mounting:12. Mount coverslip with a drop of mounting medium.

13. Seal coverslip with nail polish to prevent drying and movement undermicroscope.

14. Store in dark at -20 or 4˚C.

XIII. DOUBLE IMMUNOFLUORESCENCE-SIMULTANEOUS (IHC-P, IHC-FR, ICC) PROTOCOL

In order to be able to examine the co-distribution of two (or more) differentantigens in the same sample, a double immunofluorescence procedurecan be carried out. Primary antibodies raised in different species can beused either in parallel (in a mixture) or in a sequential way.

Preparation of slides and samples:

A. Cell lines, cytology smears, cytospin preparationsi. Coat coverslips with polyethylineimine or poly-L-lysine for 1 hr at room

temperature. ii. Rinse coverslips well with sterile H2O (3 times 5 min each). iii. Allow coverslips to dry completely and sterilize them under UV light

for at least 4 hrs. iv. Grow cells on glass coverslips or prepare cytospin or smear preparation. v. Rinse briefly in phosphate-buffered saline (PBS).

B. Frozen (cryostat) sections i. Snap frozen fresh tissue in liquid nitrogen or isopentane pre-cooled in

liquid nitrogen. Store frozen blocks at -80˚C. ii. Cut 4-8 µm thick cryostat sections and mount on superfrost or gelatin

coated slides. You can store slides at -80˚C until needed. iii. Before IF staining, warm up slides at room temperature for 30 minutes.

C. Paraffin-embedded sectionsi. Deparaffinize sections in xylene 2x5 min. ii. Hydrate with 100% ethanol 2x3 min. iii. Hydrate with 95% ethanol 1 min. iv. Rinse in distilled water and then follow procedure for fixation and

antigen retrieval as required (please see IHC protocol for formalin-fixed paraffin-embedded tissue sections for further details).

Fixation:1. Fix the samples either in ice-cold methanol, acetone (1-10 min) or in 3-4% paraformaldehyde in PBS pH 7.4 for 15 min at room temperature.

2. Wash the samples twice with ice cold PBS.

Permeabilization:If the target protein is expressed intracellularly, it is very important topermeabilize the cells. Note: acetone fixed samples do not requirepermeabilization.

3. Incubate the samples for 10 min with PBS containing 0.25% Triton X-100 (or 100 µM digitonin or 0.5% saponin). Triton X-100 is the mostpopular detergent for improving the penetration of the antibody. However, itis not appropriate for the use of membrane-associated antigens since itdestroys membranes.

4. Wash cells in PBS three times for 5 min.

Blocking and Simultaneous Incubation:5. Incubate cells with 1% BSA in PBST for 30 min to block unspecificbinding of the antibodies (alternative blocking solutions are 1% gelatin or10% serum from the species that the secondary antibody was raised in).

6. Incubate cells in the mixture of two primary antibodies (e.g. rabbitagainst human target-1 and mouse against human target-2, if the targetsare human proteins) in 1% BSA in PBST in a humidified chamber for 1 hrat room temperature or overnight at 4oC.

7. Decant the mixture solution and wash the cells three times in PBS, 5min each wash.

8. Incubate cells with the mixture of two secondary antibodies whichare raised in different species (with two different fluorochromes, i.e. Texas

Red-conjugated against rabbit and FITC-conjugated against mouse) in1% BSA for 1 hr at room temperature in dark.

9. Decant the mixture of the secondary antibody solution and wash threetimes with PBS for 5 min each in dark.

Counter staining:10. Incubate cells on 0.1-1 µg/ml Hoechst or DAPI (DNA stain) for 1 min.

11. Rinse with PBS.

Mounting:12. Mount coverslip with a drop of mounting medium.


14. Store in dark at -20˚C or 4˚C.

XIV. DOUBLE IMMUNOFLUORESCENCE-SEQUENTIAL PROTOCOL

Fixation and permeabilization, are the same as for part XIII.

Blocking and Sequential Incubation:

1. First blocking step: incubate cells with the first serum (10% serumfrom the species that the secondary antibody was raised in) for 30 min toblock unspecific binding of the antibodies (alternative blocking solutionsare 1% gelatin or 1% BSA) at room temperature.

2. Incubate cells with the first primary antibody in 1% BSA or 1% serumin PBST in a humidified chamber for 1 hr at room temperature or overnightat 4˚C depending on the concentration of the antibody and the accessibilityof the antigen.

3. Decant the first primary antibody solution and wash the cells three timesin PBS, 5 min each wash.

4. Incubate cells with first secondary antibody (labelled withFluorochrome-1) in 1% BSA in PBST for 1 hr at room temperature in dark.

5. Decant the first secondary antibody solution and wash three times withPBS for 5 min each in dark.

6. Second blocking step: incubate cells with the second serum (10%serum from the species that the secondary antibody was raised in) for 30min to block unspecific binding of the antibodies (alternative blockingsolutions are 1% gelatin or 1% BSA) at room temperature in the dark.

7. Incubate cells with the second primary antibody in 1% BSA in PBSTin a humidified chamber in the dark for 1 hr at room temperature orovernight at 4˚C depending on the concentration of the antibody and theaccessibility of the antigen.

8. Decant the second primary antibody solution and wash the cells threetimes in PBS, 5 min each wash in dark.

9. Incubate cells with second secondary antibody (labelled withFluorochrome-2) in 1% BSA for 1 hr at room temperature in dark.

10. Decant the second secondary antibody solution and wash three timeswith PBS for 5 min each in dark.

Counter staining:1. Incubate cells on 0.1-1 µg/ml Hoechst or DAPI (DNA stain) for 1 min in dark.

2. Rinse with PBS in dark.

Mounting:Mount coverslip with a drop of mounting medium.


2. Store in dark -20˚C or 4˚C.

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Fluorophores:

Nucleic acid probes:

XV. DIRECT FLOW CYTOMETRY (FACS) PROTOCOL

General Procedure: 1. Harvest, wash the cells and adjust cell suspension to a concentration of1-5x106 cells/ml in ice cold 3% BSA/PBS. Use polystyrene round-bottom12x75 mm Falcon tubes.

2. Add 0.1-10 µg/ml of the primary antibody. Dilutions, if necessary, shouldbe made in 3% BSA/PBS.

3. Incubate for at least 30 min at room temperature.

4. Wash the cells 3X by centrifugation at 400 g for 5 min and resuspendthem in ice cold PBS.

5. Dilute the fluorochrome-labeled secondary antibody in 3% BSA/PBS atthe optimal dilution (according to the manufacturer’s instructions) and thenresuspend the cells in this solution.

6. Incubate for at least 20 minutes at room temperature. This incubationmust be done in the dark.

7. Wash the cells 3x as in step 4.

8. The cells are now ready for analysis by flow cytometry. Keep the cellson ice until your scheduled time for analysis.

Remember that detection of intracellular antigens requires a cellpermeabilization step prior to staining.

XVI. INDIRECT FLOW CYTOMETRY (FACS) PROTOCOL

Procedure:Indirect labeling requires two incubation steps; one with a primary antibodythen the next one with a compatible secondary antibody. The secondary(and not the primary) antibodies have the fluorescent dye (FITC, PE, Cy5,etc.) attached. Please note that this is a general protocol and you mayneed to adjust it for your applications.

1. Harvest, wash the cells and determine the total cell number. Usepolystyrene round-bottom 12x75 mm Falcon tubes for Flowcytometry/FACS analysis. It is always useful to check the viability of thecells which should be around 95% not less than 90%.

2. Resuspend the cells approximately 1-5x107 cells/ml in ice cold 3% BSA/PBS.

3. Add 1 ml of cell suspension to each Falcon tube.

4. Add 0.1-10 µg/ml of the primary antibody. Dilutions, if necessary, shouldbe made in 3% BSA/PBS.

5. Incubate for at least 30 min at room temperature.

6. Wash the cells 3-times by centrifugation at 400 g for 5 min andresuspend them in ice cold PBS. You may need to adjust the conditions ofthe centrifugation (the force and the time) for the cell types used.

7. Dilute the fluorochrome-labeled secondary antibody in 3% BSA/PBS atthe optimal dilution (according to the manufacturer’s instructions) and thenresuspend the cells in this solution.

8. Incubate for at least 20 minutes at room temperature. This incubationmust be done in the dark.

9. Wash the cells 3-times by centrifugation at 400 g for 5 min andresuspend them in ice cold PBS.

The cells are now ready for analysis by flow cytometry. Keep the cells onice until your scheduled time for analysis. If you need to wait longer thanfor an hour, you may need to fix the cells which can preserve them for atleast several days. (This will stabilize the light scatter and inactivate mostbiohazardous agents)

10. After the last step, centrifuge the cells and remove the liquid.

11. Add 0.5 to 1.0 ml of cold 0.5% paraformaldehyde solution and vorteximmediately.

12. Store the cell suspension at 4°C in the dark.

Remember that detection of intracellular antigensrequires a cell permeabilization step prior to staining(use 0.5% saponin or acetone or Leucoperm).

DyeAbsorbanceWavelength

EmissionWavelength

Visible color

DAPI 345 455 blueHoechst 33258 345 478 blueSYTOX blue 431 480 blueHoechst 33342 343 483 blueYOYO-1 509 509 greenSYTOX green 504 533 greenTOTO 1, TO-PRO-1 509 533 greenSYTOX orange 547 570 yellowChromomycin A3 445 575 yellowMithramycin 445 575 yellowPropidium iodide 536 617 redEthidium bromide 493 620 red

DyeAbsorbanceWavelength

EmissionWavelength

Visible color

Hydroxycoumarin 325 386 bluemethoxycoumarin 360 410 blueAlexa fluor 345 442 blueaminocoumarin 350 445 blueCy2 490 510 green (dark)FAM 495 516 green (dark)Alexa fluor 488 494 517 green (light)Fluorescein FITC 495 518 green (light)Alexa fluor 430 430 545 green (light)Alexa fluor 532 530 555 green (light)HEX 535 556 green (light)Cy3 550 570 yellowTRITC 547 572 yellowAlexa fluor 546 556 573 yellowAlexa fluor 555 556 573 yellowR-phycoerythrin (PE) 480;565 578 yellowRhodamine Red-X 560 580 orangeTamara 565 580 redCy3.5 581 581 596 redRox 575 602 redAlexa fluor 568 578 603 redRed 613 480;565 613 redTexas Red 615 615 redAlexa fluor 594 590 617 redAlexa fluor 633 621 639 redAllophycocyanin 650 660 redAlexa fluor 633 650 668 redCy5 650 670 redAlexa fluor 660 663 690 redCy5.5 675 694 redTruRed 490;675 695 redAlexa fluor 680 679 702 redCy7 743 770 red



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XVII. DIRECT ELISA PROTOCOL

Buffers and Reagents:Bicarbonate/carbonate coating buffer (100 mM) - antigen or antibodyshould be diluted in coating buffer to immobilize them to the wells: 3.03 gNa2CO3, 6.0 g NaHCO3 (1000 ml distilled water) pH 9.6, PBS: 1.16 gNa2HPO4, 0.1 g KCl, 0.1 g K3PO4, 4.0 g NaCl (500 ml distilled water) pH 7.4,

Blocking solution: commonly used blocking agents are BSA, nonfat drymilk, casein, gelatin. PBS containing 1% Bovine Serum Albumin (BSA),

Wash solution: usually PBS or Tris-buffered saline (pH 7.4) with detergentsuch as 0.05% (v/v) Tween20 (TBST)

Antibody solution: primary and secondary antibody should be diluted in 1xblocking solution to prevent nonspecific bindings

Coating antigen to microplate1. Dilute the antigen to a final concentration of 20 µg/ml in PBS. Coat thewells of a PVC microtiter plate with the antigen by pipeting 50 µl of theantigen dilution per well.

2. Cover the plate with an adhesive plastic and incubate for 2 h at roomtemperature.

3. Remove the coating solution and wash the plate twice by filling the wellswith 200 µl PBS. The solutions or washes are removed by flicking the plateover a sink. The remaining drops are removed by patting the plate on a papertowel.

Blocking4. Block the remaining protein-binding sites in the coated wells by adding200 µl blocking buffer, 5% non fat dry milk/PBS, per well. Alternativeblocking reagents include BlockACE or BSA.

5. Cover the plate with an adhesive plastic and incubate for at least 2 h atroom temperature or, if more convenient, overnight at 4°C.

6. Wash the plate twice with PBS.

Incubation with the Antibody7. Add 100 µl of the antibody, diluted at the optimal concentration(according to the manufacturer’s instructions) in blocking buffer immediatelybefore use.


9. Wash the plate four times with PBS.

Detection10. Dispense 100 µl (or 50 µl) of the substrate solution per well with amultichannel pipet or a multipipet.

11. After sufficient color development (if it is necessary) add 100 µl of stopsolution to the wells.

12. Read the absorbance (optical density) of each well with a plate reader. Note: some enzyme substrates are considered hazardous (potentialcarcinogens), therefore always handle with care and wear gloves.

XVIII. INDIRECT ELISA PROTOCOL

Buffers and Reagents: (See section XVI Buffers and Reagents)

For accurate quantitative results, always compare signal of unknownsamples against those of a standard curve. Standards (duplicates ortriplicates) and blank must be run with each plate to ensure accuracy.

Incubation with Primary and Secondary antibody

1. Add 100 µl of diluted primary antibody to each well.



4. Add 100 µl of conjugated secondary antibody, diluted at the optimalconcentration (according to the manufacturer) in blocking bufferimmediately before use.

5. Cover the plate with an adhesive plastic and incubate for 1-2 h at roomtemperature.


DetectionAlthough many different types of enzymes have been used for detection,horse radish peroxidase (HRP) and alkaline phosphatase (ALP) are thetwo widely used enzymes employed in ELISA assay. It is important toconsider the fact that some biological materials have high levels ofendogenous enzyme activity (such as high ALP in alveolar cells, highperoxidase in red blood cells) and this may result in non-specific signal. Ifnecessary, perform an additional blocking treatment with Levamisol (forALP) or with 0.3% solution of H2O2 in methanol (for peroxidase).

ALP substrateFor most applications pNPP (p-Nitrophenyl-phosphate) is the most widelyused substrate.The yellow color of nitrophenol can be measured at 405nm after 15-30 min incubation at room temperature. (This reaction can bestopped by adding equal volume of 0.75 M NaOH).

HRP chromogensThe substrate for HRP is hydrogen peroxide. Cleavage of hydrogenperoxide is coupled to oxidation of a hydrogen donor which changescolour during reaction.

TMB (3,3’,5,5’-tetramethylbenzidine) add TMB solution to each well,incubate for 15-30 min, add equal volume of stopping solution (2 M H2SO4)and read the optical density at 450 nm.

OPD (o-phenylenediamine dihydrochloride. The end product is measuredat 492 nm. Be aware that the substrate is light sensitive so keep and storeit in the dark.

ABTS (2,2’-azino-di-[3-ethyl-benzothiazoline-6 sulfonic acid] diammoniumsalt. The end product is green and the optical density can be measured at416 nm.

Note: some enzyme substrates are considered hazardous (potentialcarcinogens), therefore always handle with care and wear gloves.

7. Dispense 100 µl (or 50 µl) of the substrate solution per well with amultichannel pipet or a multipipet.

8. After sufficient color development (if it is necessary) add 100 µl of stopsolution to the wells.

9. Read the absorbance (optical density) of each well with a plate reader.

XIX. ELISA USING FLUORESCENT SUBSTRATEPROTOCOL

Day 1:1. Coat with 100 µl/well of coating antibody diluted in filtered PBS.Incubate plate overnight at 4˚C, covered with plate sealer.

Day 2:2. Block plates with 200 µl/well of 4 g Block ACE powder, diluted in 100 mlof deionized water (USE a 1:4 dilution of this) for 3 hrs at RT covered withplate sealer.

3. Wash plates: PBS-T (0.05% Tween20) 250 µl/well; 3x 30 seconds.

4. Load 100 µl of standards or samples ** freshly diluted in 10% BlockACEin PBS-T overnight at 4˚C, covered with plate sealer.

Day 3:5. Incubate 100 µl/well of biotinylated reporter antibody diluted in PBS for2 hours at RT covered with plate sealer


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6. Incubate 100 µl/well of streptavidin alkaline phosphatase, 1:5000dilution in PBS for 1 hour at RT, covered with plate sealer.

7. Wash plates: TBS 250 µl/well; 3x 30 seconds.

8. Amplify signal by adding 100 µl/well AttoPhos Fluorescent substratesystem, for 5-10 min at RT. 36 mg of AttoPhos substrate should be mixedwith 60 ml of AttoPhos buffer 24 hours prior to use. Make sure well isclean-no contamination.

9. Signal measured on Fluorometer, (Victor2, Perkin Elmer); excitation:440 nm; emission: 550 nm

** • Prepare standards ahead of time.• On day of application to the plate, (day 2 in above procedures),standards are freshly diluted in 10% BSA in PBS-T from 10 ng/ml to 500pg/ml, for example.• After washing or aspirating flip plate over onto kim wipes on bench toremove excess liquid.

XX. CHROMATIN IMMUNOPRECIPITATION (ChIP)PROTOCOL

Cross-linking and Cell Harvesting 1. Start with 2 large dishes when confluent (1x107 _ 5x107 cells per dish).2. Cross-link proteins to DNA by adding formaldehyde drop-wise directly tomedia for a final concentration of 0.75% and rotate gently at roomtemperature (RT) for 10 min. 3. Rinse cells 2 x with 10 ml cold PBS. 4. Scrape cells into 5 ml cold PBS and transfer into 50 ml tube.5. Repeat with 3ml PBS. 6. Centrifuge for 5 min at 3,000 rpm, carefully aspirate off supernatant andresuspend pellet in ChIP lysis buffer (750 µl per 1x107 cells).

Sonication 7. Sonicate lysate to shear DNA to an average fragment size of 500 to1000 bp. Follow the fragment size on a 1.5% agarose gel.8. Centrifuge for 30 secs at 13,000 rpm and transfer supernatant to new tube.*9. Remove 50 µl of each sonicated sample and add to 400 µI ChIP lysis buffer.10.This sample is the input. This is used for obtaining DNA concentrationfor subsequent IP’s (see below) and as control in the final PCRs.

Determination of input DNA concentration11. Add 5 µl of proteinase K (20 mg/ml) to each input sample in ChIP lysisbuffer and heat with shaking at 65°C for 4-5 hours (or overnight) to reversecross-linking.** 12. Phenol: chloroform extract the samples and ethanol precipitate theDNA in presence of 10 µl glycogen (5 mg/ml).13. Re-dissolve the pellet in 100 µl H2O. ** 14. Transfer 5 µl of the sample in a tube containing 995 µl TE to give a200-fold dilution and read the OD260. The concentration of DNA in µg/ml isOD260 x 10,000.

Immunoprecipitation15. Use 25 µg of chromatin per IP and dilute each sample 1:10 withdilution buffer. You will need one sample for the specific antibody, and onesample for the beads-only control. 16. Add the primary antibody to all samples except the beads-only control.The amount of antibody to be added has to be determined empirically, 1-8µg per 25 µg of chromatin often works well. 17. Add 20 µl of protein A/G beads (pre-absorbed with sonicated s/ssalmon sperm DNA at 1.5 µg/20 µl beads) to all samples and IP overnightwith rotation at 4°C.18. Centrifuge the protein A/G beads for 1 min at 3,500 rpm and removethe supernatant.19. Wash beads 3 x with 1 ml wash buffer (centrifuge as above). 20. Wash beads 1 x with 1 ml final wash buffer (centrifuge as above).

Elution and reverse cross-link21. Elute DNA by adding 450 µl of elution buffer to the protein A/G beadsand rotate for 15 min at RT. 22. Spin down and transfer the supernatant into fresh tube. **23. Reverse cross-links by adding 5 µl proteinase K (20 mg/ml) to theeluates and heat with shaking at 65°C for 4-5 hours (or overnight ).**

24. Phenol:chloroform extract the samples and ethanol precipitate theDNA in presence of 10 µl glycogen (5 mg/ml).25. Resuspend pellet in 100 µl H2O and store at -20°C or proceed withdetection method (PCR, southern blot, etc). **

* Chromatin can be snap frozen in liquid nitrogen after step 8 and storedat -70˚C for up to 2 month. Avoid multiple freeze-thawing.

** The samples can be frozen and stored at -20˚C after these steps.

Solutions

ChIP Lysis Buffer 50 ml Stock solution50 mM HEPES-KOH 2.5 ml 1 M140 mM NaCl 1.4 ml 5 M1 mM EDTA pH8 0.1 ml 0.5 M1% Triton X 2.5 ml 20%0.1% Sodium Deoxycholate 0.5 ml 10%Protease Inhibs. (added fresh each time)

Dilution Buffer 50 ml Stock solution1% Triton X-100 2.5 ml 20%2 mM EDTA 0.2 ml 0.5 M150 mM NaCl 1.5 ml 5 M20 mM Tris-HCl pH 8 1 ml 1 MProtease Inhibs. (added fresh each time)

Wash Buffer 500 ml Stock solution0.1% SDS 2.5 ml 20%1% Triton X-100 25 ml 20%2 mM EDTA 2 ml 0.5 M150 mM NaCl 15 ml 5 M20 mM Tris-HCl pH8 10 ml 1 M

Final Wash Buffer 500 ml Stock solution0.1% SDS 2.5ml 20%1% Triton X-100 25ml 20%2 mM EDTA 2ml 0.5 M500 mM NaCl 50ml 5 M20 mM Tris-HCl pH8 10ml 1 M

Elution Buffer 10 ml (make fresh each time)1% SDS 0.5 ml 20%100 mM NaHCO3 1 ml 1 M

Proteinase K (Roche) Dissolve in H2O at 20 mg/ml, store at -20°C.

Protein A/G beads with s/s Sonicated Salmon Sperm 1. Dissolve Salmon Sperm DNA (Sigma) at 5 mg/ml in 100 ml H2O (4x25ml). Rotate at 4°C overnight. 2. Phenol:chloroform extract (1:1), spin for 2 min at RT at 5,000 rpm. 3. Add 30 ml EtOH and 1.2 ml KAc (3 M) to each sample, precipitate, spinas above.4. Wash with 70% EtOH, spin as above. 5. Resuspend in H2O at 10 mg/ml (rotating at RT). 6. Sonicate, big probe, for 10-20 sec (check on agarose gel). 7. Read OD260. Boil in screw-cap eppendorfs for 10-15 min then cool on ice.8. Mix equal volume of Protein A and Protein G beads. Wash 3x in PBS. 9. Remove PBS and add s/s Salmon Sperm DNA (to 1.5 µg/20 µl beads).Add PBS back up to the original volume. Incubate for 30 min with rotationat 4°C.

XXI. BUFFERS AND STOCK SOLUTIONS

Cytoskeletal bound proteins Extract Buffer:10 mM Tris, pH 7.4100 mM NaCl1 mM EDTA1 mM EGTA1 mM NaF20 mM Na4P2O7

2 mM Na3VO4


84

1% Triton X-10010% glycerol0.1% SDS0.5% deoxycholate

Soluble protein buffer:20 mM Tris-HCl, pH 7.51 mM EGTA (Ca2+ chelator)

RIPA buffer (RadioImmunoPrecipitation Assay) buffer:RIPA buffer contains the ionic detergent sodium deoxycholate as an activeconstituent and is particularly used for nuclear membrane disruption fornuclear extracts. A RIPA buffer gives low background but can denaturekinases. It can also disrupt protein-protein interactions (and may thereforebe problematic for immunoprecipitations/pull down assays).

50mM Tris HCl pH 8150 mM NaCl1% NP-40 0.5% sodium Deoxycholate 0.1% SDS

The 10% sodium deoxycholate stock solution (5 g into 50 ml) must beprotected from light.

The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O andthen add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the totalvolume to 50 ml). Store the buffer at 4˚C.

Nonidet-P40 (NP-40) buffer:20 mM Tris HCl pH 8137 mM NaCl10% glycerol1% nonidet P-402 mM EDTA

Sodium orthovanadate preparation:This needs to be done under the fume hood• Prepare a 100 mM solution in double distilled water• Set pH to 9.0 with HCl• Boil until colorless• Cool to room temperature • Set pH to 9.0 again• Boil again until colorless• Repeat this cycle until the solution remains at pH 9.0 after boiling and

cooling• Bring up to the initial volume with water• Store in aliquots at -20˚CNote: do not permit great changes in volume during boiling; put a loose lidon the container to protect from evaporation.Discard if the samples turn yellow.

TBS 10x (concentrated TBS)24.23 g Trizma HCl80.06 g NaClMix in 800 ml ultra pure water. pH to 7.6 with pure HCl.Top up to 1 L.

TBSTFor 1 L: 100 ml of TBS 10x + 900 ml ultra pure water + 1ml Tween20

Medium stripping buffer: Make fresh stripping buffer:15 g glycine1 g SDS10 ml Tween20Set the pH to 2.2 make up to 1 L with ultrapure water

Harsh stripping buffer:to be done under the fumehoodFor 100 ml: 20 ml SDS 10% 12.5 ml Tris HCl pH 6.8 0.5M67.5 ml ultra pure waterAdd 0.8ml ß-mercaptoethanol under the fumehood.

Nuclear Fractionation Protocol ReagentsBuffer A – 10 mM HEPES, 1.5 mM MgCl2, 10 mM KCl, 0.5 mM DTT,0.05% NP40 (or 0.05% Igepal or Tergitol) pH 7.9

To prepare 250 ml stock of buffer A – HEPES: 1M = 238.3 g/L, therefore 10 mM = 0.59 g/250 mlMgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250 mlKCl: 1M = 74.5 g/L, therefore 10 mM = 0.187 g/250 mlDTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250 mlNP40 = 0.05%

Buffer B – 5 mM HEPES, 1.5 mM MgCl2, 0.2 mM EDTA, 0.5 mM DTT,26% glycerol (v/v), pH 7.9

To prepare 250 ml stock of buffer B –HEPES: 1M = 238.3 g/L, therefore 5 mM = 0.295 g/250 mlMgCl2: 1M = 203.3 g/L, therefore 1.5 mM = 0.076 g/250 mlEDTA: 1M = 372.2 g/L, therefore 0.2 mM = 0.0186 g/250 mlDTT: 1M = 154.2 g/L, therefore 0.5 mM = 0.019 g/250 ml26% Glycerol (v/v) = 65 ml

4.6 M NaCl - 87.66 g/326 ml

TBS (Tris Buffered Saline) pH 7.6-7.8:

TBS 0.025% Triton X-100:

1.6% H2O2 (Hydrogen Peroxide) in TBS:

10% NS (Normal Serum) with 1% BSA (Bovine Serum Albumin,Fraction 5) in TBS:

Primary antibody made up in TBS with 1% BSA:(Example is of primary antibody used at a dilution of 1:10)

Secondary biotinylated antibody made up in TBS with 1% BSA:(Example is of secondary biotinylated antibody used at a dilution of 1:200)

ABC (Avidin-Biotin) complex in TBS:(Example is of ABC complex, each part used at a dilution of 1:100)

Bicarbonate/carbonate coating buffer (100 mM): 3.03 g Na2CO3, 6.0 gNaHCO3 (1 L distilled water) pH 9.6, PBS: 1.16 g Na2HPO4, 0.1 g KCl, 0.1 g K3PO4, 4 g NaCl (500 ml distilledwater) pH 7.4

For 1 ml: 10 µl Streptavidin

10 µl HRP (or AP)-Biotin

980 µl TBS pH 7.6-7.8

For 1 ml: 5 µl Secondary biotinylated antibody

995 µl TBS pH 7.6-7.8

For 0.1 ml: 100 µl Primary antibody

10 mg BSA

900 µl TBS pH 7.6-7.8

For 1 ml: 100 µl NS

10 mg BSA

900 µl TBS pH 7.6-7.8

For 400 ml: 6.4 ml H2O2 (GPR = 30% w/w)

393.6 ml TBS pH 7.6-7.8

For 1 litre: 250 µl Triton X-100

999.75 ml TBS pH 7.6-7.8

For 10 litres: 60.6 g TRIS HCl

13.9 g TRIS base

87.66 g NaCl10 litres Ultra pure water (H2O)


Neu

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86Ordering information: www.abcam.com | Tech support: www.abcam.com/technical

Neuroscience resources: www.abcam.com/neuroscience

Neu

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Technical help - Neuroscience signaling pathway diagrams

Homer AdenylatecyclaseGi/q

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Neurotransmission - receptor and channel signaling


Neuroscience signaling pathw

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TrkA

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- KChIP2- KCNQ 3, 5- Kv beta 2- Kv 1.2- Kv 1.3- Kv 1.4- Kv 2.2- Kv 4.2

- ARA9- ASICb , ASIC3- DDR 1, 2- ENSA- GJB1- SLC31A1- TRAR4- TRPM7- VR1- VRL1

Sodium

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Inhibitionof signaling

Cellsurvival

Growth,development &differentation

Short termplasticity

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Apoptosis

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Price and other information provided is subject to change without notice, and prices may be changed up to the time of delivery.While every effort is made to keep information provided by Abcam up to date, Abcam will not be liable to you or anyone else iferrors should occur in information provided on this website.

When your order is received Abcam will confirm receipt. However, formal acceptance of the order will only take place when thegoods are despatched. If prices should be changed between the time of receipt of an order and despatch, Abcam will contactyou in advance.

Invoices should be paid no later than 30 days after the invoice date, and customers must themselves pay any bank chargesthat are incurred in making the payment. Full payment instructions are set out on the invoice.

Receipt of productsPackaging and products should be inspected immediately upon receipt. Notification of damage, shortages or defects should begiven immediately by e-mail or fax.

Return & replacement policyQuality is important to Abcam. If a product does not perform as described on the datasheet notify us within 90 days of deliveryand our dedicated team of scientific experts will examine details of your protocol to determine whether the problem is protocolrelated or product related:

• For protocol-related problems, our experts will give you independent professional advice. • For product- related problems, a replacement or refund will be offered. • If you do not provide details of your protocol or follow the recommendations of our experts, Abcam will not issue a

replacement or refund.

If items are ordered incorrectly by the customer Abcam will consider taking them back. This will normally be subject to a 20%restocking charge on the items plus any shipping, handling & packaging costs. Requests for returns must have priorauthorization (contact [email protected]) and must be made within 7 days of receipt of the items. Items must be returned inthe same or equivalent packaging (i.e. cold and insulated) as originally despatched by Abcam.

Product use limitationsAll Abcam products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC, THERAPEUTIC ORANY IN VIVO USE IN HUMAN SUBJECTS”

Abcam will not be liable for any claims or applications not listed in the literature or the misuse of products. Abcam cannotprovide a guarantee for all applications for which a specific reagent may be used. Further, information disclosed in Abcamproduct literature should not be considered as a recommendation to use its products in violation of any patents.

Abcam shall not be liable for its failure to perform any of its obligations resulting from circumstances beyond its reasonablecontrol. Abcam will notify its customers as soon as practically possible after it becomes aware of deficiencies in a productsupplied. Any claim relating to products shall be limited to replacement or refund of original purchase price paid.

Except for personal injury or death (for which no limit applies), Abcam will not be liable (under contract, by negligence or anyother way) for any indirect or consequential loss or damage arising out of or in connection with the products or this Agreementand Abcam’s total aggregate liability for any loss or damage in respect of the products or this Agreement will not exceed theamount paid for the products.

GeneralContracts and agreements with Abcam shall in all respects be governed by and interpreted in accordance with English law.

All information correct at time of printing.


Com

ing soon

Antibody Datasheet

5HT2C Receptor antibody ab23580ALDH1A1 antibody ab23375Arg3.1 antibody ab23382BRN3A antibody ab23579Dbx1 antibody ab24605Delta-like 1 antibody ab23684Deltex antibody ab23665DLX 1 antibody ab23804DLX2 antibody ab18188DLX5 antibody ab23811DNER antibody ab23682Doublecortin (phospho S28) antibody ab23544Doublecortin (phospho S47) antibody ab23543Doublecortin (phospho S297) antibody ab23542EMX2 antibody ab23528Hippocalcin antibody ab24560KCNQ2 antibody ab22897Lingo1 antibody ab23631Mesp2 antibody ab23733Neurogenin 1 antibody ab23519Nicotinic Acetylcholine Receptoralpha 7 antibody ab23832

Antibody Datasheet

NRG1 type II antibody ab23314NRG1 type III antibody ab23248Nudel antibody ab23550PAX2 antibody ab23788PEN2 antibody ab18189Piccolo antibody ab20664PINK1 antibody ab23518PSD95 antibody ab18258Robo3 antibody ab23562Semaphorin 3c antibody ab23575Semaphorin 7a antibody ab23578Slit3 antibody ab11018Synaptojanin antibody ab19904Syntenin antibody ab19903TBR1 antibody ab23361TBR2 antibody ab23345Tbx6 antibody ab23716TPH2 antibody ab17733TRPA1 antibody ab17734USP9x antibody ab19879

Coming soon - Antibodies available soon from Abcam

Test these new antibodies today - we'd be happy to hear from you!Contact Sabrina Edmonds ([email protected]) for details.

75 newNeuroscience

antibodies addedto our catalogevery month!

92

Targ

et in

dex


Target indexProtein groupings Page14-3-3 protein . . . . . . . . . . . . . . . . . . . .14,21,405HT . . . . . . . . . . . . . . . . . . . . . . . . . . .25,50,56Acetylcholine . . . . . . . . . . . . . . . . . . . . . . .47,53Acetylcholinesterase . . . . . . . . . . . . . . . . . . .47Acrolein . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19ACTH . . . . . . . . . . . . . . . . . . . . . . . . . . . .27,50ADAM . . . . . . . . . . . . . . . . . . . . . . . . . .13,18,19Adenosine Receptor . . . . . . . . . . . . . . . . . . .55Adiponectin . . . . . . . . . . . . . . . . . . . . . . . .28,29Adrenergic Receptor . . . . . . . . . . . . . . . . . . .54Aggrecan . . . . . . . . . . . . . . . . . . . . . . . . . .11,33Agrin . . . . . . . . . . . . . . . . . . . . . . . . . . . .8,14,36AKT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14,15ALDH1A1 . . . . . . . . . . . . . . . . . . . . . . . . . . .22ALK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .37,45Als . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19,25AMIGO . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11Amisyn . . . . . . . . . . . . . . . . . . . . . . . . . . . .8,60Amitriptyline . . . . . . . . . . . . . . . . . . . . . . .53,62Amphetamine . . . . . . . . . . . . . . . . . . . . . . . . .53AMPK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18Amyloid precursor protein . . . . . . . . . . . . . . .18APH . . . . . . . . . . . . . . . . . . . . . . . . . . .13,18,22Apolipoprotein . . . . . . . . . . . . . . . . . . . . . . . .19APXL . . . . . . . . . . . . . . . . . . . . . . . . . . . . .59,63Artemin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12ASIC . . . . . . . . . . . . . . . . . . . . . . . . . . .58,59,62Ataxin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .37Attractin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28AVPR . . . . . . . . . . . . . . . . . . . . . . . . . . . .30,55Axotrophin . . . . . . . . . . . . . . . . . . . . . . . . .11,62BACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .20BAI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36BDNF . . . . . . . . . . . . . . . . . . . . . . . . . . . .12,13Bestrophin . . . . . . . . . . . . . . . . . . . . . . . . . .8,63beta Amyloid . . . . . . . . . . . . . . . . . . . . . . . . .18BMP . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12,33BNP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .50,51C3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .40,44CACNB . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43Calbindin . . . . . . . . . . . . . . . . . . . . . . . . . . . .43Calcitonin . . . . . . . . . . . . . . . . . . . . . . . . . . . .55Calcium channel L type . . . . . . . . . . . . . . . . .43Calpain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25Calpastatin . . . . . . . . . . . . . . . . . . . . . .19,40,60CaMK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .44Cannabinoid Receptor . . . . . . . . . . . . . . . . . .54CAPON . . . . . . . . . . . . . . . . . . . . . . . . . . .44,53Carboxypeptidase . . . . . . . . . . . . . . . . . . . . .28CASK . . . . . . . . . . . . . . . . . . . . . . . . . . . .40,45Caspase . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19Cathepsin . . . . . . . . . . . . . . . . . . . . . . . . .10,20CCKAR . . . . . . . . . . . . . . . . . . . . . . . .19,22,28CCKBR . . . . . . . . . . . . . . . . . . . . . . . . . . .27,55CD11 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .32

Protein groupings PageCD132 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33CD133 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33CD24 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33CD45 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .32Cdk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11Cellubrevin . . . . . . . . . . . . . . . . . . . . . . . . . . .40CGRP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .50Choline acetyltransferase . . . . . . . . . . . . .38,47CLACP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18Clathrin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .60CNPase . . . . . . . . . . . . . . . . . . . . . . . . . . . . .32CNTF . . . . . . . . . . . . . . . . . . . . . . .12,13,33,35Coilin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .38COP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45Corticoliberin . . . . . . . . . . . . . . . . . . . . . . .27,50Corticotropin Releasing Factor . . . . . .27,50,55CPEB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .40CRF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .27,32Crk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15,45CRMP . . . . . . . . . . . . . . . . . . . . . . . . . .11,25,36CSN . . . . . . . . . . . . . . . . . . . . . . . . . . .15,45,46CSPS . . . . . . . . . . . . . . . . . . . . . . . . . . . .49,61Ctip . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15,16CysLT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55Dab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14,78DARPP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .54DDR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58Dehydroepiandrosterone . . . . . . . . . . . . . . . .27DHPR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43Dispatched . . . . . . . . . . . . . . . . . . . . . . . . . . .16DLL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13Dopamine . . . . . . . . . . . . . . . . . . . . . . .49,54,61DORFIN . . . . . . . . . . . . . . . . . . . . . . . . . .22,23Doublecortin . . . . . . . . . . . . . . . . . . . . . .7,36,38Drebrin . . . . . . . . . . . . . . . . . . . . . . .8,19,35,36Dynamin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .40DYRK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11EDG . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55ELAV . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16,38EMX . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16,33Endomorphin . . . . . . . . . . . . . . . . . . . . . . . . .52ENSA . . . . . . . . . . . . . . . . . . . . . . . . . . . .26,58Ephrins . . . . . . . . . . . . . . . . . . . .11,36,45,46,59ERAB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19ERK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21Estradiol . . . . . . . . . . . . . . . . . . . . . . . . . .26,50Estrogen . . . . . . . . . . . . . . . . . . . . . . . . . .26,27Exendin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .50Fatty Acid Synthase . . . . . . . . . . . . . . . . . . . .28Fe65 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19Fetuin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8,16FMR . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16,51FOX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12,16Fyn . . . . . . . . . . . . . . . . . . . . . . . . . . . .19,20,24GABA . . . . . . . . . . . . . . . . . . . . . . .48,55,57,61

Protein groupings PageGAD . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .48Galectin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10GALR . . . . . . . . . . . . . . . . . . . . . . . . . . . .28,55GAP . . . . . . . . . . . . . . . . . . . . . . . . . . .36,38,45GCNF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16GDNF . . . . . . . . . . . . . . . . . . . . . . . . . . . .12,13Gemin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16GFAP . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6,31Ghrelin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28GHRHR . . . . . . . . . . . . . . . . . . . . . . . . . . .26,55GJB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58Gli . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16Glutamate . . . . . . . . . . . . . . . . . . . .47,48,54,61Glutamate Receptor . . . . . . . . . . . . . . . . .48,54Glycine . . . . . . . . . . . . . . . . . . . . . . . . . . .48,72Glycine Receptor . . . . . . . . . . . . . . . . . . .48,57GRM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .54GSK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21HAP40 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22Heparan sulfate proteoglycan . . . . . . . . . . . . .9HIPPI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .22HMG . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23,24Homer . . . . . . . . . . . . . . . . . . . . . . . . . . . .40,54HRH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55Humanin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19HUNK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .46Huntingtin . . . . . . . . . . . . . . . . . . . . . . . . . . .22IL6 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28,29Insulin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .28Integrin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .34Internexin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6Jagged . . . . . . . . . . . . . . . . . . . . . . . . . . .13,14Ketanserin . . . . . . . . . . . . . . . . . . . . . . . . .53,56Kv . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .59L1CAM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11Laminin . . . . . . . . . . . . . . . . . . . . . . . .6,8,24,25LANGF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12LAR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .29,40Leptin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .29LIM kinase . . . . . . . . . . . . . . . . . . . . . . . . . . .16LIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16LNX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14LPHN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55MAML . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14MAP . . . . . . . . . . . . . . . . . . . . . . .7,16,35,36,38MASS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55MC4 . . . . . . . . . . . . . . . . . . . . . . . . . . .27,29,55Melanopsin . . . . . . . . . . . . . . . . . . . . . . . . . . .63Melatonin . . . . . . . . . . . . . . . . . . . . . . .27,29,55Metabotropic Glutamate Receptor . . . . . . . . .54Mint . . . . . . . . . . . . . . . . . . . . . . . . . .8,19,40,60Morphine . . . . . . . . . . . . . . . . . . . . . . . . . . . .52MOSP . . . . . . . . . . . . . . . . . . . . . . . . . . . .10,32Munc . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .60Muscarinic Acetylcholine Receptor . . . . . .53,56

93

Target index


Protein groupings PageMyelin . . . . . . . . . . . . . . . . . . . . . . . . . .10,32,33MYRIP . . . . . . . . . . . . . . . . . . . . . . . . . .7,40,60NCAM . . . . . . . . . . . . . . . . . . . . . . . .8,16,36,37NCS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43Nestin . . . . . . . . . . . . . . . . . . . . . . . . . . .6,34,38Neurabin . . . . . . . . . . . . . . . . . . . . . . . . . . . .45Neuregulin . . . . . . . . . . . . . . . . . . . .11,13,16,33Neurofigromin . . . . . . . . . . . . . . . . . . . . . . . .45Neurofilament . . . . . . . . . . . . . . . . . . . . .6,7,37Neuroglobin . . . . . . . . . . . . . . . . . . . . . . . . . .16Neurogranin . . . . . . . . . . . . . . . . . . . . . . .44,46Neurokinin . . . . . . . . . . . . . . . . . . . . . .50,52,55Neuropeptide . . . . . . . . . . . . . . . . .29,50-52,62Neuropilin . . . . . . . . . . . . . . . . . . . . . . . . . . . .11Neuroserpin . . . . . . . . . . . . . . . . . . . . .11,25,37Neurotrophin . . . . . . . . . . . . . . . . . . . . . . .12,13Neurturin . . . . . . . . . . . . . . . . . . . . . . . . . . . .13NGF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13Nicastrin . . . . . . . . . . . . . . . . . . . . . . . . . . .8,20Nicotinic Acetylcholine Receptor . . . . . . . .57,91Nitrotyrosine . . . . . . . . . . . . . . . . . . . . . . . . . .52NMDAR . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58NMUR . . . . . . . . . . . . . . . . . . . . . . .27,29,55,62Nociception . . . . . . . . . . . . . . . . . . . . . . . .62,63Noradrenaline . . . . . . . . . . . . . . . . . . . . . . . .49NOS . . . . . . . . . . . . . . . . . . . . . . . . . . . . .53,54Notch . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13,14NPFF . . . . . . . . . . . . . . . . . . . . . . . .51,52,55,62NR2E3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16NRP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11,37NSE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39NUMB . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14Oct . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33Olfactory Receptor . . . . . . . . . . . . . . . . . . . . .63Opioid . . . . . . . . . . . . . . . . . . . . . . . . . . . .52,56Orexin . . . . . . . . . . . . . . . . . . . . . . . . .29,50,51Oxytocin . . . . . . . . . . . . . . . . . . . . . . . .30,50,51P2X . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58P2Y . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .55p35 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .37,46pan macrophage . . . . . . . . . . . . . . . . . . . . . .32Parathyroid Hormone . . . . . . . . . . . . . . . .30,55PARK7 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23Parkin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23Parvalbumin . . . . . . . . . . . . . . . . . . . . . . . . . .43PAX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16,34PBP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .47,51PDE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .63Peripherin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7Peropsin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .63PGP9.5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23PHF2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .20Phosphoserine . . . . . . . . . . . . . . . . . . . . . . . .47Phosphoserine/threonine . . . . . . . . . . . . . . . .46Phosphotyrosine . . . . . . . . . . . . . . . . . . .46,47

Protein groupings PagePhosphotyrosine phosphatase . . . . . .12,29,39PHOX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16Pin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21PINK1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23Pleiotrophin . . . . . . . . . . . . . . . . . . . . . . . . . .13Plexin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39PMP . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10,33PP2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .46PPP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .46Prealbumin . . . . . . . . . . . . . . . . . . . . . . . . . . .21Presenilin . . . . . . . . . . . . . . . . . . . . . . . . .14,20Prion protein PrP . . . . . . . . . . . . . . . . . . . . .24ProDynorphin . . . . . . . . . . . . . . . . . . . . . . . . .52Progesterone . . . . . . . . . . . . . . . . . . . . . . . . .27Prohormone Convertase . . . . . . . . . . . . . . . .29PROX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16PSD . . . . . . . . . . . . . . . . . . . . . .40,41,44,45,62Rab . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .41,60RAGE . . . . . . . . . . . . . . . . . . . . . . . . . . . .19,63RAI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16Rapsyn . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58Reelin . . . . . . . . . . . . . . . . . . . . . . . . . . . .8,9,11RIT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .63Robo . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11,37ROR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15Ryanodine . . . . . . . . . . . . . . . . . . . . . . . . . . .44S100 . . . . . . . . . . . . . . . . . . . . . . . . . . .31,32,43SAP . . . . . . . . . . . . . . . . . . . . . . . . . . .36,41,45SCP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .16Serotonin . . . . . . . . . . . . . . . . . . . . . . . . . .50,56SHANK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45SHC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19SIAH . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14,23SIRP . . . . . . . . . . . . . . . . . . . . . . . . . . . .8,17,41Slit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11,17SNAP . . . . . . . . . . . . . . . . . . . . . . . . . .41,60,61SOX . . . . . . . . . . . . . . . . . . . . . . . . . . .17,34,39Spinesin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10SPON . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13Sprouty . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17Stathmin . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12STEP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .39Substance P . . . . . . . . . . . . . . . . . . . . . . .51,52Survivin . . . . . . . . . . . . . . . . . . . . . . . . . . .33,39Synapsin . . . . . . . . . . . . . . . . . . . . . . .41,60,61Synaptobrevin . . . . . . . . . . . . . . . . . . . . . .41,61Synaptophysin . . . . . . . . . . . . . . . . . . .41,42,61Synaptotagmin . . . . . . . . . . . . . . . . . . . .9,42,60SynCAM . . . . . . . . . . . . . . . . . . . . . . . . .9,17,42Syndecan . . . . . . . . . . . . . . . . . . . . . . . . . . . .42SynGAP . . . . . . . . . . . . . . . . . . . . . . . . . . . . .45Synphilin . . . . . . . . . . . . . . . . . . . . . . . . . . . .23Syntaxin . . . . . . . . . . . . . . . . . . . . . . . . . .42,61Syntrophin . . . . . . . . . . . . . . . . . . . . . . . . . .9,58Synuclein . . . . . . . . . . . . . . . . . . . . . . .20,23,24

Protein groupings PageTau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .21,22Taurine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .48Tenascin . . . . . . . . . . . . . . . . . . . . . . . . . . .9,17Thrombospondin . . . . . . . . . . . . . . . . . . . . . . .7Thyroid Hormone . . . . . . . . . . . . . . . . . . . . . .30TRAF . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17TRAR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .58Trk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .13TRPM . . . . . . . . . . . . . . . . . . . . . . . . . . . .58,62Tryptamine . . . . . . . . . . . . . . . . . . . . . . . .48,49Tryptophan . . . . . . . . . . . . . . . . . . . . . . . . . . .49TSH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .30Tubulin . . . . . . . . . . . . . . . . . .7,25,33,34,36,38Tyramine . . . . . . . . . . . . . . . . . . . . . . . . . .48,49Ubiquitin . . . . . . . . . . . . . . . . . . . . . . . .20,22,25UCP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .29VAMP . . . . . . . . . . . . . . . . . . . . . . . . . . . .42,61Vanilloid . . . . . . . . . . . . . . . . . . . . . . . .58,62,63Vasopressin . . . . . . . . . . . . . . . . . . . . . . .30,51VCAM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17VCP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .24VGLUT . . . . . . . . . . . . . . . . . . . . . . . . . . .53,61Vimentin . . . . . . . . . . . . . . . . . . . . . . . .34,35,37VIP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .51VRL . . . . . . . . . . . . . . . . . . . . . . . . . . .51,58,63Zic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .17

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