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Perkin Elmer HydroGel slide and the SCHOTT alternative

IntroductionSCHOTT can offer an alternative to the PE slide: SCHOTT Nexterion Slide H.

PE microarray slide PE Product Code

Equivalent SCHOTT slide

SCHOTT Product Code

HydroGel coated slides-Single pad (25 per box) no barcode

6050017 Nexterion Slide H bar-coded 25 slides per pack

1070936

Further Nexterion Slide H information & protocols are available onhttp://www.schott.com/nexterion/english/http://www.schott.com/nexterion/english/download/protokol_slide_h.pdf

Comparison PE HydroGel Slide and SCHOTT Slide H performancesPerkin Elmer HydroGel coated slides

Nexterion Slide H Comments

Applications Protein microarrays Protein microarraysSmall molecules (peptides, sugars, etc.)

Both PE HydroGel and Slide H may be used for arraying proteins without the need to modify the probes.

Coating type and thickness

3-Dimensional surfaceThe HydroGel gel is 4 microns thick when dry and 20-30µm thick when hydrated.

3-Dimensional surfaceSlide H coating is 10 -60 nm.

The PE surface is very fragile when hydrated meaning that it can be very easily damaged by contact printers. The Slide H surface is much thinner, and less prone to damage.However due the reduced thickness, customers will need to re calibrate the printing (Z axis) height when they change to Slide H

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Pre printing treatment step

The PE HydroGel slides have a protective storage agent applied to the surface after manufacture. This layer has to be removed in a four-step procedure. .

Slide H may be used immediately from the packaging (after bringing the package up to room temperature. Slide H must be stored at -20'C ). No pre-treatment is required

Chemical composition

The PE HydroGel slide uses an acrylamide-based polymer coating bound to a glass slide.

Slide H is based on a type of polymer specifically chosen for its extremely low non-specific binding.

Typically the non-specific binding on Slide H is much lower than on acrylamide-based polymers.

Binding capacity

Proteins are irreversibly immobilized on the PE slide within the porous HydroGel substrate, through interactions with the matrix. The recommended blocking for the PE is a "physical" blocking method using PBST + 1%BSA. Non-specific binding of serum proteins can still occur on the blocked HydroGel coating, leading to a higher background.

Slide H has a high binding capacity as a result of having NHS-ester active groups for covalently binding bio molecules. After arraying, the residual NHS ester groups can be efficiently chemically deactivated by treating with 50 mM ethanolamine in buffer.

Slide H will typically show a higher signal to background ratios that the PE HydroGel.

Protocols For details see below For details see below The PE HydroGel and Slide H protocols are very different, and we would only recommend using the SCHOTT Slide H protocols with the SCHOTT slides.

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Comparison PE HydroGel Slide and SCHOTT Slide H ProtocolsStep PE HydroGel SCHOTT Slide HStorage (of unprinted slides)

+20°C, sealed Short term: (< 2 week) +20°C, dry and sealedLong term: sealed, -20°C

Pre-printing wash procedure

Required (to remove storage agent)• 3 x (briefly) dH2O • 3 x 10 min dH2O• 6 x (briefly) dH2O• spin dry: 1000-1500 RCF, 5 min• 20 min, 40°C

Not required

Print buffer • Phosphate / borate buffer systems• Glycerol up to 40%• PH stability 5-9

Recommended printing buffer (for contact printing)150 mM Phosphate, pH 8.5, 5 % Glycerol, 0,1 mg/ml BSA, 0,01%Sarcosyl

Immobilization 8-16 h at 30°C, approximately 65-70% air humidity (saturated sodium chloride solution)• 20 sek PBST (0.5%)• 3 x 30 min PBST

Over night at 20°C, approximately 65-70% air humidity (saturated sodium chloride solution)

Blocking / Deactivation

Optional• PBST with 1% BSA, 1h, RT• 3 x (briefly) PBST

Required (to deactivate remaining NHS ester groups)PBST (0.01%) with 50 mM EthanolaminePBST, 5 mindH2O, 1 min

Target Incubation Recommended dilution buffer for target / anatlyte:• PBSTWashes:• (Briefly) PBST (0.5%)• 3 x 10 min PBST (0.1%)

Recommended dilution buffer for target / anatlyte:• PBST (0.05%)Washes:• PBST (1%), 2 min• PBST (0.5%), 2 min• PBST (0.1%), 2 min• 2 x (briefly) dH2Ospin dry (e.g. 1000 RPM, 2 min)

Date: 12. Februar 2007