BIOXTRA...n g e p n l e s BIOXTRA GE Healthcare Life Sciences and VWR bring protein workflows together in a symphony of solutions. New products! MabSelect PrismA Protein preparation2

Isolation& cloning

Sampleprep

Purification

Cellculture

Analysis

BIOXTRA™

GE Healthcare Life Sciences and VWR bring protein workflows together in a symphony of solutions.

New products!

MabSelect™ PrismA

Protein preparation syringe filter

2

Isolation and cloningHow the calculator works . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 04

Sanger sequencing: past successes and current applications . . . . . . 04

Promotional products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 05

Cell cultureHeat inactivation—Are you wasting your time . . . . . . . . . . . . . . . . . . . . 07


Sample prepImprove lab efficiency through better filtrations . . . . . . . . . . . . . . . . . . 09

What’s in your filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 09

Save time on HPLC Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10


PurificationInterpreting protein binding capacity for chromatography resins. . . . .16

Protect your column . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Use of affinity chromatography for antibody purification . . . . . . . . . 14


AnalysisReproducible protein enrichment: a benchmark analysis . . . . . . . . . 18

Tips for a successful immunoprecipitation . . . . . . . . . . . . . . . . . . . . . . . . 18


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Table of contents

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Resources such as handbooks, posters, and selection guides

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Established brands to support your research needs

Purification AnalysisSample prepCell cultureIsolation and cloning

ÄKTA™

Chromatography columns, resins and systems

Key products:• HiTrap™ columns• HisTrap™ columns• PD-10 desalting

columns• HiLoad™ columns• Superdex™ Increase

GL columns• Ni Sepharose™

resin• MabSelect™ PrismA

resin• Protein G

Sepharose resin• Capto™ Q resin• Capto S ImpAct resin• Capto ImpRes resin

Key products:• NC and PVDF

membranes• Amersham ECL™ gels• CyDye™ labelling kits• ECL detection

reagents• Rainbow™ markers• Hyperfilm™• Amersham

QuickStain• PlusOne reagents• Electrophoresis and

transfer units

Key products:• Puradisc syringe

filters• SPARTAN™ syringe

filters• Whatman GD/X™

syringe filters• Mini-UniPrep™ filter

vials• 934-AH™ RTU• GF/C™ RTU• Polycap SPF• RC membrane• Vacu-Guard• Benchkote™• Custom designed

filtration

Key products:• ExoProStar™ S• FTA™ cards• GenomiPhi™• Nucleon™• PuReTaq and Hot

Start RTG™ PCR beads

• SC GenomiPhi • TempliPhi™

Key products:• DMEM• RPMI• FBS• Reagents and buffers• Process water

Products for cell separation and isolation:• Percoll™• Ficoll™

Amersham™ Systems, membranes, films and reagents

Whatman™

Laboratory filtration products

illustra™

Nucleic acid sample prep and storage

HyClone™

Cell culture media and sera

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How to use the Percoll calculatorUndiluted Percoll products can be diluted directly to make a final working solution of known density, by the following procedure. In a measuring cylinder, add 1.5 M NaCl or 2.5 M sucrose to 1/10 of the final desired volume (e.g., 10 mL for 100 mL of working solution). To this, add the required volume of Percoll/Percoll PLUS (undiluted), calculated using the following formula. Make up to the final volume with distilled water.

The calculator is based on the formula: V0 = V (ρ - 0.1ρ10 - 0.9 / ρ0 - 1)

Where:

V0 = volume of Percoll (undiluted; mL)

V = volume of final working solution (mL)

ρ = desired density of the final solution (g/mL)

ρ0 = density of Percoll (undiluted) (g/mL; see Certificate of Analysis for exact density)

ρ10 = density of diluting medium (g/mL)

Examples of common diluting medium:

• For cell work: 1.5 M NaCl = 1.058 g/mL (minor differences for other salts)

• For subcellular particles or viruses: 2.5 M sucrose = 1.315 g/mL

The formula is useful for achieving densities that will be very close to the actual density desired. However, slight variations in volumes and densities of diluting media will affect final density. For determining highly accurate densities, measure the final density of Percoll solutions using a densitometer or refractometer.

Sanger sequencing: past successes and current applicationsBy Rebecca Evans, Global Marketing Manager, Genomics and Diagnostic Solutions, GE Healthcare

It’s easy to think of Sanger sequencing as a technology of the past in this era of ever-increasing speed and scale. But Frederick Sanger’s DNA sequencing approach remains relevant even in today’s sequencing landscape—40 years after its introduction.

Let’s explore Sanger’s dideoxy chain termination method and how it continues to help scientists today.

What is Sanger sequencing?

Sanger sequencing is a method for determining the nucleotide sequence of DNA molecules. Developed by two-time Nobel laureate Frederick Sanger and his colleagues in 1977, it enabled an international collaboration of scientists to deliver the first human genome sequence.

By the mid-1970s, we had long known the basic structure of DNA and mechanism of transcription. But sequencing technology was in its infancy, and studying more genes and diseases in less time required a more efficient method to read DNA sequences.

The Sanger method relies on using dideoxy chain terminating nucleotides to produce sequence fragments of graduated lengths, each equal to the position of the base complementing the terminating nucleotide. Reading the DNA sequence involves visualizing these fragments and identifying the terminating nucleotides in order of fragment length (Fig. 1).

6 READ MORE HERE

In the 1980s, developments in fluorescent detection, polymerase chain reaction (PCR), and electrophoresis improved the ease-of-use, speed, and sensitivity of Sanger sequencing. This automation-friendly approach led scientists to consider sequencing the human genome a real possibility—something only dreamt of a decade earlier!

Commercial Sanger sequencing equipment, such as the MegaBACE 4000 (Amersham International, now part of GE Healthcare) and PRISM 3700 (Applied Biosystems), were powerhouses of the Human Genome Project. Started in 1990, the project completed its objective—sequencing the human genome— in 2003, two years ahead of schedule.

Isolation and cloning

Percoll Calculator

Workflow for the Sanger method of DNA sequencing. Each dideoxynucleotide (ddNTP) is labeled using one of four different fluorescent dyes, enabling analysis on a single lane and automation.

1. Starting with enough DNA sample, add a primer that binds to your region of interest.

2. Add your labeled and unlabeled nucleotides. A small fraction (around 1%) of terminating fluorescently-labeled dideoxynucleotides (ddNTPs) enables fragment visualization. Each fluorescently labeled ddNTP carries a different fluorophore (red, yellow, blue, and pink).

3. Synthesize to obtain your labeled fragments of varying lengths.

4. Separate the fragments by capillary electrophoresis with single-base accuracy and find your original sequence.

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illustra PuReTaq Ready-To-Go™ PCR beadsPremixed, predispensed, single-dose reactions optimized for performing standard PCR amplifications.

• Save time: simply add template DNA solution and primers, and cycle.

• More reproducible results: less risk of pipetting errors and contamination.

• High-quality PuReTaq DNA polymerase and high-purity reagents: robust performance and the lowest possible levels of contaminating DNA.

• Long-term ambient-temperature stable: no freezer space required; less energy consumption for shipping and storage.

• Verified for real-time polymearase chain reaction (PCR).

• Available in 96 well plates and Hot Start Mix.

illustra ExoProStar reagentExoProStar 1-Step PCR and sequence reaction clean-up reagent provides a simple, single-step method for removing unincorporated primers and nucleotides prior to downstream analysis. illustra Exonuclease I and Alkaline Phosphatase enzymes are combined in a single, stable mix requiring just one pipetting step in a 30 min protocol to establish the clean-up reaction.

ExoProStar S is an enzymatic PCR clean-up technology with Exonuclease I and Shrimp Alkaline Phosphatase to remove unincorporated primers and dNTPs. For maximum flexibility it comes in two separate tubes; just two simple pipetting steps are needed to prepare the reaction.

ExoProStar S is easy to automate and provides complete heat inactivation of the enzymes within 10 min in a fast 15 min protocol. It is scalable for different reaction sizes and easy to automate.

illustra TempliPhi amplification kitsPrepare DNA sequencing templates from circular DNA for cycle sequencing, cloning, and transformation in 4 to 6 hours.

• Use simple protocol to reduce time, labor, and quantity of consumables needed for template preparation–workflow enables easy automation.

• Use amplified DNA directly for cycle sequencing without purification.

• Amplify DNA from bacterial or M13 liquid cultures, colonies, plaques, glycerol stocks, or purified circular (plasmid or M13) DNA.

Isolation& cloning

Cellculture


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What is single-cell sequencing: challenges and applicationsBy Andrew Gane, G&Dx Solutions Strategy & Technology Manager at GE Healthcare

Drilling down to the single-cell level provides insights into complex biological systems and diseases. Learn about single-cell sequencing, its applications and challenges.

Single-cell sequencing: an overview

Standard bulk methods of cell analysis use many thousands of cells. When we analyze that data, we’re effectively averaging out any small cell-to-cell variances and concentrating on features or data points that rise above the noise.

It’s easy to assume that all cells of the same timepoint from the same sample are, well, the same. But that’s far from the case. Cell populations are heterogeneous: studying single-cell samples is crucial to understanding these complex biological systems. Drilling down to the single-cell level allows us to understand the effects on and contributions by individual cells within their environment.

What is single-cell genomics?

Single-cell genomics applies standard analytical techniques, including sequencing and microarrays, to the individual cell, utilizing advanced techniques for selecting and handing individual cells and maximizing the raw material (DNA, RNA, proteins) held within. Single-cell genomics has numerous applications in both basic research and clinical settings.

6 READ MORE HERE

Binding of target mRNA occurs through the pairing of the polyadenylated RNA tail found on the 3’ end of mRNA to the covalently bound oligo(dT) groups on the surface of the Sera-Mag Oligo(dT) Magnetic Particles. This binding is easily accomplished using standard hybridization conditions

A. Total RNA Sample

C. Pairing of polyadenylated tail D. Apply magnetic field for mRNA Separation

B. Mix Oligo(dT) with RNA smaple

The power of attraction for nucleic acid isolationSera-Mag™ Oligold(T)-Coated Magnetic Particles easily isolates and extracts your valuable mRNA from a variety of sources and enables you to perform such applications as RT-PCR cDNA library construction, cDNA microarrays, affinity purification, primer extension and subtractive hybridization.

• Convenient isolation and extraction of mRNA from a variety of sources for downstream applications such as reverse transcription-PCR (RT-PCR), complementary DNA (cDNA) library construction, cDNA microarrays, affinity purification, primer extension, and subtractive hybridization.

• Covalently bound oligo(dT)14 prevents leaching from the particle surface.

• Very high, specific poly A+ binding capacity ensures maximum extraction of messenger RNA (mRNA).

• Fast reaction kinetics increases throughput and precision, and also enables faster movement through viscous solutions.

Encapsulation

Magnetite

Core

Isolation& cloning

Cellculture



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Description and origin Filtration En

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Specialty fetal bovine serumSuper Low IgG FBS, US 40 nm ≤ 10 ≤ 10 • • • • • • • • • • • ≤ 5

Charcoal/Dextran Treated FBS, US Triple 100 nm ≤ 10 ≤ 20 • • • • • • • • • • • •

Dialyzed FBS, US Triple 100 nm ≤ 10 ≤ 10 • • • • • • • • • • • •

ES Cell Screened FBS, US 40 nm ≤ 10 ≤ 10 • • • • • • • • • • • •

Tetracycline Screened FBS, US 40 nm ≤ 10 ≤ 10 • • • • • • • • • • • •

Insect Cell Screened FBS, US 40 nm ≤ 10 ≤ 10 • • • • • • • • • • • •

Human Mesenchymal SC Screened FBS

40 nm ≤ 10 ≤ 10• • • • • • • • • •

• •

• = Tested. For typical results, request a certificate of analysis or go to www.gelifesciences.com/HyClone

HyClone specialty serum

Screened FBS productsPrescreened from 40 nm filtered Defined FBS, our specialty serum products meet the needs of many specific cell culture applications

HyClone tetracycline screened Fetal Bovine Serum (FBS), U.S. origin

• FBS with undetectable levels of tetracycline.

Human Mesenchymal Stem Cell Screened FBS

• Designed for human mesenchymal stem cells

ES Cell Screened FBS

• Optimized for the culture of murine embryonic stem cells.

Insect Cell Screened FBS

• To optimize performance of insect cells.

Heat inactivation—are you wasting your time?Heat inactivation is a process commonly applied to bovine serum products used for mammalian cell culture. This process involves raising the temperature of the serum to 56°C for a period of 30 min. As a serum supplier, we are often questioned if this process is necessary and what the purpose of heat inactivation of cell culture serum might be. In this article, we describe reasons for heat inactivation and how these reasons may no longer be valid for the majority of cell culture applications. Studies were completed to compare the

growth performance of serum with a number of commonly used cell lines. Results demonstrate no advantage to subjecting serum to heat activation, and in many cases it exhibited a negative effect on cell growth. It is our conclusion that heat inactivation is not necessary for the majority of mammalian cell culture applications.

6 READ MORE HERE

Cell culture


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Media

• Classical media

• Serum free media

• Insect media

• Chemically defined media

• Stem cell media

Sera

• Fetal bovine serum

• Specialty fetal bovine serum

• Bovine calf serum

• Species-specific serum

• Engineered serum

Reagents and supplements

• Cell dissociation reagents

• Antibiotics and selection reagents

• Cryopreservation media

• Feed supplements

• Amino acids

• Vitamins

• Glucose

• Cholesterol and lipids

Buffers and process liquids

• Balanced salt solutions

• Buffered saline solutions

• Cleaning and sanitation solutions

• Water (molecular biology grade, cell culture grade, WFI quality)

HyClone cell culture productsWith a proven record as the trusted global manufacturer of cell culture media, serum, and process liquids, we provide the products that facilitate cell culture based research and biopharmaceutical production.

• High quality FBS and cell culture reagents maximize cell growth.

• Animal sera with certified traceability offer reliability.

• Consistent, secure supply network ensures high level assurance of stock.

• True pool processing increases bottle-to-bottle consistency.

• Innovative serum processing techniques to meet your demands.

Media filtrationPolycap TC 36 Capsule Filter, 0.2/0.1 μm, Sterile, with SB Inlet and Outlet (1 pc)

Whatman Polycap TC filtration capsules from GE Healthcare feature dual layer polyethersulfone (PES) membranes and provide efficient filtration for aqueous solutions.

• 100% integrity-tested during manufacturing.

• Polypropylene housing thermally fused to eliminate surfactants and mold releasing agents that could affect analysis.

• Manufactured according to ISO9001:2008 QMS standards.

Isolation& cloning

Cellculture


Polypropylene support materials

Pleated membrane cartridge

Polypropylene housing

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Improve lab efficiency through better filtrationDo you consider particle retention, loading capacity, and liquid flow rate when choosing a filter or device?

Perhaps there is a better filter out there for your application. Or perhaps your analysis might be easier, quicker, or produce results that are more consistent if you switched your filter to a different grade.

Here are three key characteristics to consider when identifying the right filter:

1. Particle retention—For cellulose and glass microfiber papers, it is expressed as a “nominal retention rating”, and quoted at 98% efficiency to allow for secondary filtration effects. For membrane filters with defined pore sizes, it is an absolute retention rating.

2. Loading capacity—Filters with the highest loading capacities are chemically treated and are more expensive than their untreated counterparts. Treatment might also interfere with analysis. This can happen either through chemical interaction with the sample or by increasing the time to results due to a slower flow rate than that of an untreated filter. By knowing the weight of filtrate that you want to retain on the filter, you can choose a filter that will safely accommodate your needs without the downsides of a filter that is more complex than is needed.

3. Liquid flow rate—The flow rate describes the speed at which a liquid flows through the filter. In practice, this is dependent on several factors that will often be specific to the solid/liquid being filtered. But, for comparison purposes, a typical water flow rate is measured and provided for each grade under gravity and normalized to a certain diameter.

6 READ MORE HERE

Grade 1 qualitative filter papersA widely used standard grade filter paper for routine applications with medium retention and flow rate. It covers a wide range of laboratory applications and is frequently used for clarifying liquids.

Sample preparation

What’s in your filter?Chemical compatibility in HPLC samples

Efficient high-performance liquid chromatography (HPLC) analysis requires removal of particulates that could interfere with accurate detection of the analyte of interest. Filtration prior to HPLC serves to reduce impurities but risks introducing new contaminants in the form of extractables.

Awareness and selection of syringe filter components based on their chemical compatibility to solvents reduces the risk of new contaminants interfering with HPLC analyses.

What are common methods of sample preparation?

Accurate HPLC analyses require suitable sample preparation to maximize purity and ensure the analyte concentration is within the range of detection. Common methods for sample preparation include:

• Dilution

• Centrifugation

• Filtration

Dilution or centrifugation of individual samples might be sufficient to minimize insoluble particles for HPLC. Sample filtration using a filtration device adds an extra layer of security, because the membrane physically segregates insoluble particles to help prevent irregular chromatogram peaks. Syringe and syringeless filters are a convenient option to process multiple samples for autosampling.

However, filtration devices themselves can be a source of extractables with the potential to influence data quality, dependent on their compatibility with the solvents used in sample preparation.

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Save time in HPLC prepSample filtration protects your HPLC instrument and column while preserving data quality.

If you analyze large numbers of samples using HPLC, sample preparation can take up a lot of your time. Filtering samples before HPLC can help avoid frit clogging.

Try a stacked syringe filter

Syringe filtration often involves aspirating the sample, fitting a particle filter, filtering into an autosampler vial, capping, and finally transferring the vial to an autosampler. You might repeat this process dozens of time a day, depending on your circumstances.

If you have difficult-to-filter samples, you might find that high particulate samples can

take more time to filter. To help with this, stacked filter devices have multiple layers of filtration media, starting with larger pore sizes and going down to the desired pore size.

This approach traps large particles first and successively traps smaller particles (see picture). The device does not get clogged as easily as devices with a single membrane, making filtration faster and easier.

Go syringeless

If your samples are reasonably easy to filter, a syringeless filter option simplifies the process greatly.

In a syringeless filter, the filter membrane, pre-filtration chamber, post- filtration storage vial, and cap are all part of one device

(see picture). This design streamlines HPLC sample prep and minimizes the number of consumables. Filtration can be performed three times faster than with syringe filters.

Using a syringeless filter means that you only need to add the sample to the outer chamber, place the plunger, and push. The inner storage vial holds your filtered sample ready for analysis, so it can go directly into your autosampler. Construction can be either polypropylene or glass and the vial can be either clear or amber colored depending on the requirements of your sample.

Septum

Cap

Locking ring

Plunger

Chamber

Filtering liquid

Membrane

Liquid to be filtered(containing particulates)

Sampleflow

Layers 3GF/F: 0.7 µm

Layers 4Choice of membranes and pore sizes

Whatman GD/X stacked syringe filter Mini-UniPrep syringeless filters

Mini-UniPrep syringeless HPLC filters

Whatman Mini-UniPrep syringeless filters integrate an autosampler vial, filtration membrane, plunger, and cap/septa into one consumable product. They are built for fast and easy HPLC/UHPLC sample preparation.

• 0.2 µm and 0.45 µm pore sizes available to meet sample requirements.

• Housing options: amber to prevent photodegradation of light-sensitive samples, or translucent for easy visual inspection.

• Compatible with most major autosamplers for high throughput analysis.

• All-in-one filtration device for quick and cost-effective sample processing.

Whatman GD/X Syringe Filters—Prefilter, Non-Sterile

Whatman GD/X syringe filters from GE are specifically designed for filtration of viscous or otherwise hard-to-filter samples with high solids content.

• Glass fiber prefiltration stack for filtering larger sample volumes with less back pressure build-up.

• High loading capacity for samples with high solids content.

• Processes three to seven times more sample volume than filters without prefilter.

Featured products to keep your column clean

Isolation& cloning

Cellculture


6 READ MORE HERE


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Chemical compatibility of membranes and housingsSolvent ANP CA CN PC PE GMF NYL PP DpPP PES PTFE† PVDF RCAcetic acid, 5% R LR R R R R R R R R R RAcetic acid, glacial R NR NR R LR R R R R R NRAcetone R NR NR NR R R R R R NR R NR RAcetonitrile R NR NR R R R R NR R R RAmmonia, 6N NR NR NR LR LR R R R R R LR LRAmyl acetate LR NR NR NR R R R R R LR R LR RAmyl alcohol R LR LR R R R R NR R R RBenzene* R R R NR R R LR NR NR R R R RBenzyl alcohol* R LR LR LR R R LR R R NR R R RBoric acid R R R R R R LR R R R R RButyl alcohol R R R R R R R R R R R R RButyl chloride* R NR NR NR R RCarbon tetrachloride* R NR R LR R R LR NR NR NR R R RChloroform* R NR R NR R R NR LR LR NR R R RChlorobenzene* R LR NR R NR LR NR R R RCitric acid R LR R R R R RCresol* NR R R NR NR NR NR R NR RCyclohexanone R NR NR R NR R R NR R R RCyclohexane* R NR NR R R R NR NR NR NR R R RDiethyl acetamide NR NR R R R R R NR RDimethyl formamide LR NR NR R R R R NR R NR LRDioxane R NR NR NR R R R R R LR R LR RDMSO LR NR NR NR R R R R R NR R LR LREthanol R R NR R R R R R R R R R REthers* R LR LR R R R R NR NR R R LR REthyl acetate R NR NR NR R R R R R NR R NR REthylene glycol R LR LR R R R R R R R R R RFormaldehyde LR LR R R R R R LR LR R R R LRFreon TF* R R R R R R NR NR NR R R RFormic acid LR LR R NR R R R R R LRHexane R R R R R R R R R R R R RHydrochloric acid, conc* NR NR NR NR NR R NR LR LR R R R NRHydrofluoric acid* NR NR NR NR LR LR R R NRIsobutyl alcohol R LR LR R R R R R R R R RIsopropyl alcohol R R LR R R R R R R RMethanol R R NR R R R R R R R R R RMethyl ethyl ketone R LR NR NR R R R R R NR R NR RMethylene chloride* R NR LR R NR LR LR NR R R RNitric acid, conc* NR NR LR NR R NR NR NR NR R R NRNitric acid, 6N* LR LR R NR LR LR LR R R LRNitrobenzene* LR NR NR NR R R LR R R NR R R RPentane* R R R R R R R NR NR R R R RPerchloro ethylene* R R R R LR NR NR NR R R RPhenol 0.5% LR LR R R NR R R NR R R RPyridine R NR NR NR R R LR R R NR R NR RSodium hydroxide, 6N NR NR NR NR NR NR LR R R R R NR NRSulfuric acid, conc* NR NR NR NR NR R NR NR NR NR R NR NRTetrahydrofuran* R NR NR R R LR LR NR R R RToluene* R LR R NR R R LR LR LR NR R R RTrichloroethane* R NR LR NR R R LR LR LR NR R R RTrichloroethylene* R R R NR LR LR NR R R RWater R R R R R R R R R R R R RXylene* R R R R LR LR LR LR R R R

R = Resistant; LR = Limited Resistance; NR = Not Recommended* Short-term resistance of housingThe above data is to be used as a guide only. Testing prior to application is recommended.† Membrane may need pre-wetting with isopropanol/methanol if filtering a polar liquid

Material abbreviations: ANP – AnoporeCA – Cellulose AcetateCN – Cellulose NitrateDpPP – Polypropylene Depth Filter

GMF – Glass MicrofiberNYL – NylonPC – PolycarbonatePE – PolyesterPES – Polyethersulfone

PP – PolypropylenePTFE – PolytetrafluoroethylenePVDF – Polyvinylidene DifluorideRC – Regenerated Cellulose

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Whatman 250 yearhttps://tinyurl.com/ ya5ul43a

We are thinking Pink in OctoberOctober is breast cancer awareness month and GE will be supporting the Breast Cancer Research Foundation (BCRF).

GE Healthcare’s Whatman laboratory filtration products are used in laboratories supporting critical work in areas from cancer research to drug discovery and development. Learn more at:gelifesciences.com/ThinkPink2019

To find out more about the work of the BCRF check the link:bcrf.org/researchers

What’s the difference: Autovial https://tinyurl.com/ yy2g6ojt

What’s the difference: MiniUni-Prephttps://tinyurl.com/ y563vlm9

VIDEOs

Whatman: 250 years of innovationFor over 250 years, Whatman has been a trusted name for quality filtration products. We live up to that history every day with innovative solutions for the present and future.

Whatman environmentalhttps://tinyurl.com/ y38dqz5o

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Sample preparation with the Protein Prep syringe filter for ÄKTA systemsProtein Prep syringe filters are ready-to-use with polycarbonate housing and a regenerated cellulose membrane that is low protein binding and broadly compatible with common solvents. Syringe filtration has been shown to reduce debris residue in the column that could otherwise affect performance and column life. In addition, the Protein Prep syringe filter is lot certified for low levels of extractable particles that might otherwise interfere with chromatograms.

Protein prep syringe filter for ÄKTA systems

• 13 mm or 30 mm diameter.

• 0.2 µm or 0.45 µm pore size.

Tips for choosing the right filter

• Use 13 mm diameter filter for sample volumes < 10 mL.

• Use 0.2 µm pore size filter if the particle size of the chromatography resin is < 30 µm.

Protect your columns and your peaksGood practice in liquid chromatography (LC) is to filter samples prior to injection onto the column. Sample filtration prevents unwanted particulates from entering the column. This is important because particulates can reduce column lifetime, increase run time, and distort peak shape. In addition to affecting the quality of your results, particulates might clog the column inlet, causing increased back pressure and premature ending of chromatography runs. Adjusting protocols to extend usable lifetime can minimize the frequency of column replacement while maintaining data quality throughout.

Although filtration prior to chromatography is common practice, the factors that affect ideal filter selection often are not considered. The ‘right’ filter depends on the method that the sample is being prepared for, the chemical properties of the solvent being used, and the physical and chemical properties of the sample itself.

The new Protein Prep syringe filter for protein work has been tested for: 1) the impact of filtration on column cleanliness, 2) the recovery using regenerated cellulose as a preparation medium and 3) the filter quality to the fidelity of results.

Our regenerated cellulose is batch tested to ensure low levels of extractable compounds that might otherwise interfere with analyses.

Percent recovery of BSA samples filtered with regenerated cellulose membranes

Pore sizeRecovery at 1 mg/mL (%) SD

Recovery at 0.5 mg/mL (%) SD

0.2 µm 98 0.6 97 0.3

0.45 µm 99 0.7 99 0.9

SD = Standard deviation, N = 3

Hold-up volumes of water in filtration devices containing regenerated cellulose membranes. Results before and after purge

Pore size

Hold-up volume (μmL) Final hold up volume (μmL)

Average SD Average SD

0.2 µm 135 36 9.3 0.9

0.45 µm 135 31 9.2 1.1

SD = Standard deviation, N = 10

Chromatograms of water filtered through 30 mm 0.2 µm regenerated cellulose membrane syringe filters and subjected to HPLC analysis. Top panel shows results at 254 nm; bottom panel shows results at 215 nm.

The clean-in-place (CIP) peak after purification of the unfiltered (orange) and filtered (green and blue) mAb samples.

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Purification


14

Use of affinity chromatography for antibody purificationHow does antibody purification work?Antibodies are members of a family of molecules, the immunoglobulins. Polyclonal antibodies, monoclonal antibodies (mAb), and antibody fragments are usually purified by affinity chromatography. Resins containing an immobilized ligand (e.g., protein A, protein G, or protein L) are used to capture antibodies and antibody fragments (right). IgG, which is by far the most common immunoglobulin, is commonly purified with protein G and protein A, both of which have a strong affinity to the Fc region of IgG. Protein L has a strong affinity to the variable region of kappa light chains.

What do antibody purification schemes look like? Antibody purification protocols typically are challenged by two factors. The first is specifically capturing as many antibodies as possible in the first step as well as controlling the degradation of the sample. The second is removing the remaining impurities and minimizing the aggregate content. Below you will find suitable protocols to choose from. The 2-step protocol is the recommended best choice for research use. The 3-step protocol considers upscaling or process development needs. Size exclusion chromatography (SEC) is not used as a final step to remove aggregates, fragment or other impurities due to the limitations of sample volume. Instead, a combination of IEX steps is used.

6 READ MORE HERE

Capture

Isolate, concentrate, stabilize

Intermediate purification

Remove bulk impurities

Polishing

Remove final contaminants

Moderate to high purity High yield

Step 1

Very high purity

Good balance purity/yield

Step 1

Step 2

Highest purity Moderate yield

Step 1

Step 2

Step 3

Protein purification Protein purification methods can be organized as a single or multi-step chromatographic strategies to optimize the required purity and yield. A strategy can comprise capture (C), intermediate purification (iP), and polishing steps (P).

• Capture or affinity chromatography is where the target protein of interest is isolated, concentrated, and stabilized.

• Intermediate chromatography is used to remove contaminants with techniques including ion exchange chromatography (IEX), cation exchange chromatography (CIEX) and hydrophobic interaction chromatography (HIC).

• Polishing is utilized to remove the most difficult impurities and aggregates of the target protein to increase purity.

Depending on your downstream analysis technique, a single or multi-step chromatographic process can be followed by sample desalting, buffer exchange, and concentration.

Isolation& cloning

Cellculture



15

MabSelect PrismAHiTrap columns are pre-packed with MabSelect PrismA protein A chromatography resin. The affinity resin in the packed column has been improved with an optimized high-flow agarose base matrix and a genetically engineered protein ligand, allowing efficient cleaning between monoclonal antibody purification runs. Improved resin helps meet future demands in monoclonal antibody purification, including processing of many bispecific antibodies.

• Enhanced dynamic binding capacity (DBC) compared with other protein A resins.

• Excellent alkaline stability enables efficient cleaning and sanitization using 0.5 to 1.0 M NaOH.

• Convenient HiTrap format for easy connection to a syringe, peristaltic pump, or chromatography systems such as an AKTA™ system for process optimization.

DBC of MabSelect PrismA, Protein A Sepharose High Performance, and rProtein A Sepharose Fast Flow for a polyclonal human IgG after multiple cycles with 1 M NaOH included in each cycle.

Size exclusion chromatography columnsSize exclusion chromatography (SEC) is typically used as a last polishing step in a protein purification protocol. We offer a wide range of gel filtration or SEC media (resins).

Next generation size exclusion chromatography columns packed with Superdex Increase and Superose Increase resinsThese columns are designed for high resolution in small-scale preparative purification and analysis for sample volumes up to 500 µL.

They deliver improved performance compared to predecessor columns:

• Reduced run time with maintained resolution.

• Higher resolution for improved protein purity.

• Higher lot-to-lot consistency.

Superose 6 IncreaseLarger proteins and protein complexes† Fractionation range Mr ~ 5000 to 5 000 000

Superdex 75 IncreaseRecombinant tagged proteins Fractionation range Mr ~ 3000 to 70 000

Superdex 200 IncreasemAb and other antibodies Fractionation range Mr ~ 10 000 to 600 000

Superdex 30 IncreasePeptides and other small biomoleculesFractionation range Mr ~ 100 to 7000

HiTrap MabSelect PrismAHiTrap rProtein A FFHiTrap Protein A HP

0

20

40

60

80

100

120

0 5 10 15 20 25 30

Rem

aini

ng re

lativ

e D

BC (%

)

Cycle

Isolation& cloning

Cellculture


16

Interpreting protein binding capacities for chromatography resinsComparing protein binding capacities of resins from different suppliers is not straightforward, because different methods may have been used and these methods are not always stated. However, binding capacity is a critical parameter, because it determines how much resin is needed to purify a certain amount of protein.

The capacities that suppliers specify for their chromatography resin may be based on different: (1) modes of measurement (static or dynamic), (2) experimental conditions (pH, salt/conductivity, protein identity and concentration) and (3) units of measure (i.e., capacity per milliliter wet chromatography medium or per gram dry medium). When inquiring about these conditions, it is worthwhile to ask the vendor whether the binding capacity was determined under static or dynamic conditions.

The static binding capacity (SBC, also called total protein capacity) is normally measured in batch mode in a beaker. SBC is usually reported as the maximum amount of protein bound to a chromatography resin at given solvent and protein concentration

conditions. Dynamic binding capacity (DBC) is the binding capacity under operating conditions (i.e., in a packed affinity chromatography column during sample application). The DBC of a chromatography resin is the amount of target protein that binds under given flow conditions before a significant breakthrough of unbound protein occurs. A breakthrough curve is generated by graphing the amount of protein loaded

versus the percent breakthrough. The DBC can be determined on the breakthrough curve at a loss of, for example, 10% protein (referred to as the QB10 value, see picture).

6 READ MORE HERE

25

20

15

10

5

00 20 30 40 50 60

Brea

kthr

ough

(%)

mg protein load/medium volume (mg/mL)

Dynamic binding capacity determination breakthrough curve

QB10 = 48 mg/mL

Speeding up run time for viscous samplesViscous samples, such as E. coli extract, can increase back pressure on the column and force you to reduce the flow rate, resulting in longer load times. Utilizing a robust, modern, and rigid resin—such as Capto Q ImpRes—reduces sample run time

significantly by allowing high flow rate with low column back pressure, as seen in the example below.

HiTrap Capto ImpRes contains Capto ImpRes high-flow agarose resin, which has

much better pressure/flow properties than Sepharose High Performance resin. (Capto ImpRes resin has maximum flow rate of 300 cm/h vs 150 cm/h for Sepharose High Performance.) In this example, the IEX total run time is 30% shorter.

Example with 50 mL E. coli lysate run in cold room

Equi

libra

tion

2.5

min

Equi

libra

tion

2.5

min

Wash and gradient elution

15 min

Wash and gradient elution

15 min

HiTrap Capto ImpRes 1 mLSample loading – 1.9 mL/min

26 min

HiTrap Q HP 1 mLSample loading – 1.1 mL/min

45 min

Total time 43.5 min

Total time 62.5 min

Isolation& cloning

Cellculture



17

Longer shelf life18 months shelf life on ECL Select and Prime products

StabilityECL products are stable and

stored at room temp

Western blotting detection Amersham ECL detection reagents based on horseradish peroxidase (HRP)-conjugated secondary antibodies have become the most commonly used detection method for Western blotting. It is a sensitive detection method, where the light emission is proportional to protein quantity.

Minute quantities of proteins can be detected and quantitated.

Amersham Western blotting membranesGE Healthcare Life Sciences offers a broad selection of nitrocellulose (NC) and polyvinylidene difluoride (PVDF) Western blotting membranes, with pore size ranges to suit your application requirements.

• Optimized for chemiluminescent and fluorescent detection.

• Excellent protein binding capacity over a wide size range.

• New larger pack sizes reduce your price per blot by up to 30%.

Selection guide—Amersham Western blotting membranesGE Healthcare offers scientists excellent membranes in a selection of rolls, sheets, and sandwiches.

Analysis

Small proteins and peptides < M r 10 000


Proteins in a wide range of molecular weights




High-abundant protein







Low-abundant protein





Chemiluminescence

Stripping and reprobing

Fluorescence

Amersham Protran 0.1 NC



Amersham Protran Supported 0.2 NC

Amersham Hybond P 0.2 PVDF


Amersham Protran Premium 0.2 NC

Amersham Protran Supported 0.45 NC



Amersham Protran Premium 0.45 NC

Amersham Hybond LFP 0.2 PVDF

Comparison of

0.1 µm

0.2 µm

0.2 µm

0.2 µm

0.2 µm

0.2 µm

2.5

2.5

2.5

0.3

0.3

0.3

0.03

0.03

0.03

0.8

0.8

0.8

0.09

0.09

0.09

0.00

90.

009

µg Pore size

Pore size

Pore size

µg

1.4

1.4

1.4

0.15

0.15

0.15

0.01

60.

016

0.5

0.5

0.5

0.05

0.05

0.05

µg

0.2 µm

0.45 µm

0.45 µm

0.45 µm

0.45 µm

0.45 µm

Comparison of Amersham Protran Supported and Amersham Hybond P membranes.

Comparison of Amersham Protran Premium and Amersham Hybond LFP membranes.

18

Analyzeprecipitated

antigen

Antigen sample

Antigen sample

Antibody

Antibody

Immunecomplex

Immunecomplex

Separate complex

Separate complex

Discard Discard

Pre-immobilizedantibody approach

Free antibodyapproach

Bead

edsu

ppor

tBe

aded

supp

ort

Bead

edsu

ppor

tBe

aded

supp

ort

Bead

edsu

ppor

tBe

aded

supp

ort

Bead

edsu

ppor

t

Tips for successful immunoprecipitationFor immunoprecipitation (IP), there are four main components that you can control to get superior results: protocol, resin, format, and antibody.

Choose direct or indirect method as basic protocol shown in the picture. In the direct method (pre-immobilized antibody) the antibodies are attached to the beads in advance. The direct method gives you better control of the antibody binding and is the only way if you wish to immobilize the antibodies covalently to the beads. This way, you can make sure that the antibodies will not elute and interfere with the subsequent analysis. In the Indirect method (free antibody) antibodies are incubated with the antigen before beads are added. The indirect method is very easy to use, has fewer steps, and can be used if the antibodies do not interfere with following steps.

There are two well-suited protein ligands for binding IgG type of antibodies: Protein A and Protein G. Both bind strongly to the constant region of the antibody, meaning that they will not interfere with the antibody-antigen binding. Protein A and Protein G bind to slightly different parts of the antibody, which will impact binding to antibodies from different species.

Finally you need to decide on a suitable format, depending on the number of samples, laboratory equipment, and your personal preferences.

6 READ MORE HERE

Reproducible protein enrichment: a benchmark analysisTo demonstrate efficiency and reproducibility, Protein A Mag Sepharose was evaluated in GE Healthcare’s laboratories by running nine replicates. As a comparison, a parallel experiment was set up with Dynabeads™ Protein A (Invitrogen Corporation).

Human transferrin labeled with Cy™ 5 minimal dye, was enriched from a background of non-labeled E. coli proteins using a polyclonal rabbit anti-human transferrin antibody.

For both media, the cross-link protocol was followed, and each medium was treated according to the instructions delivered with the corresponding product.

Eluted fractions were analyzed with SDS-PAGE (ExcelGel™), poststained with Deep Purple total protein stain, and scanned in Ettan™ DIGE Imager.

Protein A Mag Sepharose displayed a significantly higher recovery compared with Dynabeads Protein A, a recovery average of 53% and 20%, respectively (Fig 2). The purity was more or less the same with a purity average of 52% for Protein A Mag Sepharose and 50% for Dynabeads Protein A respectively.

6 READ MORE HERE

Protein A Mag SepharoseProtein A Mag Sepharose is designed to simplify enrichment of target proteins by immunoprecipitation or pull-down applications. It consists of magnetic beads based on Sepharose with native protein A as ligand.

• Visible and dense Sepharose-based magnetic beads with protein A ligand, easy to spot and collect bound target protein.

• Non-adherent beads eliminate smearing effects and aggregate formation; can be used without detergents.

• Simple capture of target protein in small or large sample volumes (low microliter to high milliliter scale).

Isolation& cloning

Cellculture


A

B

Replicate no.

Replicate no.



19

VACU-GUARD VACU-GUARD disc filters available in 50 and 60 mm formats.

• Excellent for protecting vacuum pump systems from solvent vapor or gaseous contaminants and aqueous aerosols.

• Designed for in-line use with stepped barb connections for 10 to 12 mm i.d. hose.

• Available with choice of chemical trap: activated carbon, molecular sieve, or desiccant.

Benchkote surface protectorGE Healthcare’s Whatman Benchkote surface protector is an absorbent lab paper that protects surfaces against hazardous spills.

• Strong: Benchkote surface protector is strong and tear resistant when wet or dry.

• Ease-of-use: Smooth white surface can be written on with ink or pencil and lies flat.

• Clean: Suitable for saturation with disinfectant to protect benches where pathogens and other bacteria are present.

• Disposable: Benchkote paper can be incinerated after use.

Lab essential

Strips, pH range 0 to 14, pH indicators and test papers• Instant pH readings

• Accurate for a wide range of routine pH testing

• Inexpensive

• Convenient and portable for field use

Lens cleaning tissue

High-quality Whatman lens cleaning tissue is chemically pure and free from silicones and other additives.

• Soft texture will not damage lenses or optical surfaces.

• Chemically pure tissue is free from silicones and other additives.

• High absorbency leads to increased safety upon removal of surface moisture and grease.

• Very strong and leaves no fibers.

Analysis

20

Isolation and cloningItem description Ordering code Ordering quantity

Density gradient media

Percoll 89428-522P 250 mL

Ficoll-Paque PLUS 95038-170P 6 × 500 mL

Cloning and amplification

illustra ExoProStar 71002-802P 500 rxn

illustra TempliPhi 500 amplification kit 89131-612P 500 rxn

illustra TempliPhi sequence resolver kit 89131-618P 200 rxn

illustra Single Cell GenomiPhi 10146-198P 100 rxn

illustra GenomiPhi HY kit 95040-348P 100 rxn

illustra RNAspin mini 95017-489P 20 preps

illustra RNAspin 96 Mini kit 95017-499P 4 × 96 preps

Ready-To-Go (RTG) GenomiPhi V3 kit 89233-918P 96 rxn

illustra Ready-To-Go RAPD analysis kit 95040-340P 100 rxn and 6 primers

First-Strand cDNA synthesis kit 95040-330P 55 rxn

Ready-To-Go (RTG) You-Prime First-Strand beads 95040-334P 50 rxn

Ready-To-Go (RTG) RT-PCR beads, 0.2 mL 89497-128P 0.2 mL tubes

illustra GFX 96 PCR purify kit 95026-726P 96-well plates

illustra GFX PCR DNA and gel band purification kit 95026-730P 250 preps

Sera-Mag beads

Sera-Mag Magnetic Olido(dt) particles, 1 µm, 1% solids, 0.05% Na azide 10204-656P 1 mL

Sera-Mag Magnetic carboxylate modified particles, 1 µm, 5% solids, 0.05% Na azide 10204-628P 15 mL

Nucleic acid blotting

Nytran™ SuPerCharge (SPC) blotting membrane 300 mm × 3 m 28151-318P 1 roll

Nytran (N) Nylon blotting membrane 0.2 µm, 300 mm × 3 m 28151-400P 1 roll

Nytran (N) Nylon blotting membrane 0.45 µm, 300 mm × 3 m 28151-360P 1 roll

Hybond™-N+ positively charged nylon membrane 300 cm × 3 m 95038-400P 1 roll

Hybond-N neutral charged nylon membrane 20 cm × 3 m 95038-382P 1 roll

Plant genomics

illustra Nucleon PhytoPure 0.1 g 95026-702P 50 preps × 0.1 g

illustra Nucleon PhytoPure 1.0 g 95026-704P 50 preps × 1.0 g

Whatman FTA plantsaver 80077-200P 100/pk

UP TO 30% OFF LIST

21

Sample preparationItem description Ordering code Ordering quantity

Syringe filters

Whatman Protein prep syringe filter for ÄKTA systems 30 mm / 0.2 µm RC NEW 76291-876P 150/pk

Whatman Protein prep syringe filter for ÄKTA systems 13 mm / 0.45 µm RC NEW 76291-880P 150/pk

Whatman GD/X prefilter 25 mm / 0.2 µm Nylon 28138-154P 150 µk

Whatman GD/X prefilter 25 mm / 0.45 µm Nylon 28138-156P 150 µk

Whatman GD/X prefilter 25 mm / 0.2 µm PTFE 28138-200P 1500 µL

Whatman GD/X prefilter 25 mm / 0.45 µm PTFE 28138-202P 1500 µL

Whatman GD/XP prefilter 25 mm / 0.45 µm PVDF 28137-994P 1500 µL

Syringeless syringe filters

Whatman MiniUni-Prep 0.2 µm Nylon 14224-976P 100/pk

Whatman MiniUni-Prep 0.45 µm Nylon 28137-754P 100/pk

Whatman MiniUni-Prep Amber 0.45 µm PTFE 83009-802P 100/pk

Whatman MiniUni-Prep G2, Glass 0.2 µm RC 10036-036P 100/pk

Whatman MiniUni-Prep G2, Glass 0.45 µm PVDF 89234-968P 100/pk

Membranes and papers

Whatman RC55 0.45 µm, 47 mm round membrane 97010-098P 100/pk

Whatman Nylon 0.2 µm, 47 mm round membrane 28159-723P 100/pk

Whatman Grade 1, 12.5 cm 28450-128P 100/pk

Whatman Grade 41, 15 cm 28478-080P 100/pk

Venting, vacuum and in-line filters

Whatman VACU-GUARD 60 mm, 0.45 mm PTFE 28137-737P 10/pk

Whatman Polydisc In-line filter 50 mm, 0.2 µm PTFE 28137-852P 10/pk

Whatman PolyVENT 500 mm, 0.2 µm PTFE 28137-656P 1/pk

Whatman Polycap 150 Capsules Sterile Tissue Culture 0.8 / 0.2 µm PES 83009-782P 1/pk

Whatman Polycap 36 Capsules Sterile Tissue Culture 0.8 / 0.2 µm PES 83009-778P 1/pk

Lab Eessentials—pH strips, bench protection and lens paper

Whatman Strips, pH range 0 to 14, pH indicator and test papers 6 × 85 mm 10034-824P 100/pk

Whatman Benchkote Sheets 46 × 57 cm 10035-340P 100/pk

Whatman Benchkote Plus 60 × 50 m 52857-997P 1 reel

Whatman 105 10 × 15 cm lense tissue paper 97002-542P 25/pk

Cell cultureItem description Ordering code Ordering quantity

Serums

HyClone Fetal bovine serum USDA tested 89133-098P 500 mL

HyClone Bovine growth serum (US) 82006-908P 500 mL

HyClone Fetal bovine serum ES screened (US) 43300-310P 500 mL

HyClone Fetal bovine serum insect cell screen 82007-420P 500 mL

HyClone Fetal bovine serum hMSc screened (US) 95059-626P 500 mL

HyClone Fetal bovine serum tetracyline screened 82007-422P 500 mL

Waters and buffers

HyClone molecular biology grade water 82007-336P 1000 mL

HyClone cell culture grade water 82007-330P 1000 mL

HyClone Water For Injection (WFI) Quality Water 16750-108P 6 × 1000 mL

HyClone 1X Phosphate buffered saline without calcium and magnesium 16777-251P 500 mL

HyClone 1X phosphate buffered saline without calcium and magnesium 16750-122P 6 × 1000 mL

HyClone 1X Dulbecco’s phosphate buffered saline without calcium and magnesium (DPBS) 16750-076P 6 × 500 mL

HyClone1X Dulbecco’s phosphate buffered saline with calcium and magnesium 16777-259P 1000 mL

Classic culture media

HyClone RPMI 1640 media with L-glutamine 16750-070P 6 × 500 mL

HyClone RPMI 1640 media with L-glutamine 16777-145P 6 × 500 mL

HyClone RPMI 1640 without L-glutamine cell culture media 16750-084P 6 × 500 mL

HyClone RPMI 1640 with HEPES and L-glutamine cell culture media 16777-367P 500 mL

HyClone DMEM /high glucose, L-glutamine and sodium pyruvate 16777-200P 500 mL

HyClone DMEM /high glucose and L-glutamine 16777-129P 500 mL

HyClone DMEM/F12 1:1 media with L-glutamine and HEPES 16777-133P 500 mL

UP TO 80% OFF LIST

UP TO 40% OFF LIST

22

PurificationItem description Ordering code Ordering quantityAffinity chromatographyHisTrap HP 95055-912P 5 × 1 mLHisTrap Excel 89233-412P 5 × 1 mLHiTrap MabSelect PrismA NEW 76237-728P 5 × 1 mLMabSelect PrismA Resin NEW 76237-732P 200 mLIon exchange, hydrophobic and multimodal chromatographyHiTrap Capto IEX selection kit 97068-042P 7 × 1 mLHiTrap Capto Q 95056-218P 5 × 1 mLHiTrap Capto S 95056-214P 5 × 1 mLHiScreen Capto Phenyl (high sub) 97067-836P 1 × 4.7mLHiTrap Capto Core 700 89230-834P 5 × 1 mLHiTrap Capto adhere 95056-158P 5 × 5 mLHiTrap Capto adhere ImpRes 89425-694P 5 × 1 mLHiTrap HIC selection kit 95055-860P 7 × 1 mLSize exclusion chromatographySuperose 6 Increase 10/300 GL 10192-228P 1 eaSuperdex 75 Increase 10/300 GL 75799-300P 1 eaSuperdex 200 Increase 10/300 GL 89497-272P 1 eaSuperdex 30 Increase 10/300 GL 76015-026P 1 eaMagnetic beadsSera-Mag SpeedBead Protein A/G particles, 1 µm, 1% solids, 0.05% Na azide 10204-616P 5 mLSpeedBead Magnetic Neutravavidin Coated particles, 1 µm, 1% solids 10204-760P 5 mLSera-Mag SpeedBead Blocked Streptavidin particles, 1 µm, 1% solids 10204-622P 5 mLMag Rack 6: 6 positions for 1.5 mL microcentrifuge tubes 89129-096P 1 eaMagRack Maxi: 1 position for 14 mL conical, 1 position for 50 mL conical 97060-912P 1 ea

AnalysisItem description Ordering code Ordering quantity

ImmunoprecipitationMagRack 6: 6 positions for 1.5 mL microcentrifuge tubes 89129-096P 1 eaMagRack Maxi: 1 position for 14 mL conical, 1 position for 50mL conical 97060-912P 1 eaSera-Mag SpeedBead protein A/G particles, 1 µm, 1% solids 10204-616P 5 mLProtein G Mag Sepharose 89129-076P 500 µLSera-Mag SpeedBead blocked streptavidin particles, 1 µm, 1% solids 10204-626P 5 mLGel electrophoresisDTT 95017-128P 5 gTEMED 95017-114P 25 mLAmersham ECL Rainbow molecular weight marker, 50 gels 95040-114P 250 µLTransferAmersham Semi-Dry Transfer Unit, TE 77 , 21 × 26 cm 95044-802P 1 eaAmersham Protran Nitrocellulose membrane, 0.2 µm, 30 cm × 4 m 10120-004P 1 rollAmersham Protran Nitrocellulose membrane, 0.45 µm, 30 cm × 4 m 10063-173P 1 rollAmersham Protran Premium Nitrocellulose membrane, 0.45 µm, 30 cm × 4 m 10120-006P 1 rollAmersham Hybond PVDF membrane, 0.2 µm, 26 cm × 4 m 10120-022P 1 rollAmersham Hybond PVDF membrane, 0.45 µm, 30 cm × 4 m 10061-492P 1 rollWhatman Grade 3 mm chromatography paper, 20 × 20 cm 21427-364P 100 pkBlotting and detectionAmersham ECL Western blotting reagent, 4000 cm2 95038-564P 500 mLAmersham ECL Prime Western blotting reagent, 1000 cm2 89168-782P 100 mLAmersham ECL Select Western blotting reagent, 1000 cm2 89233-310P 100 mLAmersham ECL Start Western blotting reagent, 2000 cm2 10662-310P 200 mLAmersham ECL Mouse IgG, HRP-linked whole Ab (from sheep) 95017-332P 1 mLAmersham ECL Rabbit IgG, HRP-linked whole Ab (from donkey) 95017-556P 1 mLAmersham Hyperfilm ECL 8 × 10 in 95017-661P 100 shtsEnzyme immunoassayscAMP Biotrak EIA (non-acetylation protocol) competitive enzyme immunoassay system 95019-098P 96 rxncGMP Biotrack EIA competitive enzyme immunoassay system 95019-102P 96 rxn

UP TO 20% OFF LIST

UP TO 30% OFF LIST

23

Color my lab world

GE, the GE Monogram, 934-AH, ÄKTA, Amersham, Benchkote, Capto, Cy, CyDye, ECL, Ettan, ExcelGel, ExoProStar, Ficoll, FTA, GD/X, GenomiPhi, GF/C, HiLoad, HisTrap, HiTrap, Hybond, HyClone, Hyperfilm, illustra, MabSelect, Mini-UniPrep, Nytran, Percoll, Rainbow, Ready-To-Go, RTG, Sepharose, Sera-Mag, SPARTAN, Superdex, TempliPhi, and Whatman are trademarks of General Electric Company.

Dynabeads is a trademark of Thermo Fisher Scientific. Nucleon is a trademark of Gen-Probe Life Sciences Ltd. All other third-party trademarks are the property of their respective owners.

*The Polymerase Chain Reaction (PCR) is covered by patents owned by Roche Molecular Systems and F Hoffmann-La Roche Ltd. A license to use the PCR process for certain research and development activities accompanies the purchase of certain reagents from licensed suppliers such as GE Healthcare and affiliates when used in conjunction with an authorized thermal cycler.

Phi 29 DNA polymerase and its use for DNA synthesis is covered by US patent numbers 5,854,033, and 5,576,204.

Ready to Go RT-PCR Beads: Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,789,224, 5,618,711, and 6,127,155. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

© 2019 General Electric Company.

All offers in this issue are valid until December 15th 2019

All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request.

Contact your local GE Healthcare representative for the most current information.

Every effort will be made to give reasonable notice of price changes but we reserve the right to change prices without further notice. All prices exclude value added or sales tax and may be subject to change without notice. These offers cannot be used in conjunction with any previously arranged purchase agreement.

KA8717220719BR

0719 Lit. No. 070063 Promo Code 4353

Documents

BIOXTRA...n g e p n l e s BIOXTRA GE Healthcare Life Sciences and VWR bring protein workflows together in a symphony of solutions. New products! MabSelect PrismA Protein preparation2