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  Perkin Elmer HydroGel slide and the SCHOTT alternative
Introduction
SCHOTT can offer an alternative to the PE slide: SCHOTT Nexterion Slide H.

PE microarray slide          PE Product Equivalent        SCHOTT SCHOTT
                             Code       slide                    Product Code

HydroGel coated slides- 6050017            Nexterion Slide H bar-    1070936
Single pad (25 per box) no                 coded 25 slides per
barcode                                    pack



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

Comparison PE HydroGel Slide and SCHOTT Slide H performances
             Perkin Elmer HydroGel      Nexterion Slide H         Comments
             coated slides
Applications Protein microarrays        Protein microarrays      Both PE HydroGel
                                        Small molecules          and Slide H may
                                        (peptides, sugars, etc.) be used for
                                                                 arraying proteins
                                                                 without the need
                                                                 to modify the
                                                                 probes.

Coating type 3-Dimensional surface      3-Dimensional surface     The PE surface is
and          The HydroGel gel is 4      Slide H coating is 10 -   very fragile when
thickness    microns thick when dry     60 nm.                    hydrated meaning
             and 20-30µm thick when                               that it can be very
             hydrated.                                            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
For immediate release


Pre printing   The PE HydroGel slides        Slide H may be used
treatment      have a protective storage     immediately from the
step           agent applied to the          packaging (after
               surface after                 bringing the package
               manufacture. This layer       up to room
               has to be removed in a        temperature. Slide H
               four-step procedure. .        must be stored at -20'C
                                             ). No pre-treatment is
                                             required
Chemical    The PE HydroGel slide            Slide H is based on a     Typically the non-
composition uses an acrylamide-              type of polymer           specific binding on
            based polymer coating            specifically chosen for   Slide H is much
            bound to a glass slide.          its extremely low non-    lower than on
                                             specific binding.         acrylamide-based
                                                                       polymers.
Binding        Proteins are irreversibly     Slide H has a high        Slide H will
capacity       immobilized on the PE         binding capacity as a     typically show a
               slide within the porous       result of having NHS-     higher signal to
               HydroGel substrate,           ester active groups for   background ratios
               through interactions with     covalently binding bio    that the PE
               the matrix. The               molecules. After          HydroGel.
               recommended blocking          arraying, the residual
               for the PE is a "physical"    NHS ester groups can
               blocking method using         be efficiently
               PBST + 1%BSA. Non-            chemically deactivated
               specific binding of serum     by treating with 50 mM
               proteins can still occur on   ethanolamine in buffer.
               the blocked HydroGel
               coating, leading to a
               higher background.
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.
For immediate release


Comparison PE HydroGel Slide and SCHOTT Slide H Protocols
Step                  PE HydroGel                   SCHOTT Slide H
Storage (of unprinted +20°C, sealed                 Short term: (< 2 week) +20°C,
slides)                                             dry and sealed
                                                    Long term: sealed, -20°C
Pre-printing wash       Required (to remove storage Not required
procedure               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
Print buffer            •      Phosphate / borate   Recommended printing buffer
                        buffer systems              (for contact printing)
                        •      Glycerol up to 40%   150 mM Phosphate, pH 8.5, 5 %
                        •      PH stability 5-9     Glycerol, 0,1 mg/ml BSA, 0,01%
                                                     Sarcosyl
Immobilization          8-16 h at 30°C,              Over night at 20°C,
                        approximately 65-70% air     approximately 65-70% air
                        humidity (saturated sodium   humidity (saturated sodium
                        chloride solution)           chloride solution)
                        •      20 sek PBST (0.5%)
                        •      3 x 30 min PBST
Blocking /              Optional                      Required (to deactivate
Deactivation            •      PBST with 1% BSA,      remaining NHS ester groups)
                        1h, RT                        PBST (0.01%) with 50 mM
                        •      3 x (briefly) PBST     Ethanolamine
                                                      PBST, 5 min
                                                      dH2O, 1 min
Target Incubation       Recommended dilution          Recommended dilution buffer
                        buffer for target / anatlyte: for target / anatlyte:
                        •       PBST                  •       PBST (0.05%)
                        Washes:                       Washes:
                        •       (Briefly) PBST (0.5%) •       PBST (1%), 2 min
                        •       3 x 10 min PBST       •       PBST (0.5%), 2 min
                        (0.1%)                        •       PBST (0.1%), 2 min
                                                      •       2 x (briefly) dH2O
                                                      spin dry (e.g. 1000 RPM, 2 min)

Date: 12. Februar 2007

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Perkin Elmer HydroGel slide and the SCHOTT alternative (Slide H)

  • 1. For immediate release Perkin Elmer HydroGel slide and the SCHOTT alternative Introduction SCHOTT can offer an alternative to the PE slide: SCHOTT Nexterion Slide H. PE microarray slide PE Product Equivalent SCHOTT SCHOTT Code slide Product Code HydroGel coated slides- 6050017 Nexterion Slide H bar- 1070936 Single pad (25 per box) no coded 25 slides per barcode pack Further Nexterion Slide H information & protocols are available on http://www.schott.com/nexterion/english/ http://www.schott.com/nexterion/english/download/protokol_slide_h.pdf Comparison PE HydroGel Slide and SCHOTT Slide H performances Perkin Elmer HydroGel Nexterion Slide H Comments coated slides Applications Protein microarrays Protein microarrays Both PE HydroGel Small molecules and Slide H may (peptides, sugars, etc.) be used for arraying proteins without the need to modify the probes. Coating type 3-Dimensional surface 3-Dimensional surface The PE surface is and The HydroGel gel is 4 Slide H coating is 10 - very fragile when thickness microns thick when dry 60 nm. hydrated meaning and 20-30µm thick when that it can be very hydrated. 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
  • 2. For immediate release Pre printing The PE HydroGel slides Slide H may be used treatment have a protective storage immediately from the step agent applied to the packaging (after surface after bringing the package manufacture. This layer up to room has to be removed in a temperature. Slide H four-step procedure. . must be stored at -20'C ). No pre-treatment is required Chemical The PE HydroGel slide Slide H is based on a Typically the non- composition uses an acrylamide- type of polymer specific binding on based polymer coating specifically chosen for Slide H is much bound to a glass slide. its extremely low non- lower than on specific binding. acrylamide-based polymers. Binding Proteins are irreversibly Slide H has a high Slide H will capacity immobilized on the PE binding capacity as a typically show a slide within the porous result of having NHS- higher signal to HydroGel substrate, ester active groups for background ratios through interactions with covalently binding bio that the PE the matrix. The molecules. After HydroGel. recommended blocking arraying, the residual for the PE is a "physical" NHS ester groups can blocking method using be efficiently PBST + 1%BSA. Non- chemically deactivated specific binding of serum by treating with 50 mM proteins can still occur on ethanolamine in buffer. the blocked HydroGel coating, leading to a higher background. 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.
  • 3. For immediate release Comparison PE HydroGel Slide and SCHOTT Slide H Protocols Step PE HydroGel SCHOTT Slide H Storage (of unprinted +20°C, sealed Short term: (< 2 week) +20°C, slides) dry and sealed Long term: sealed, -20°C Pre-printing wash Required (to remove storage Not required procedure 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 Print buffer • Phosphate / borate Recommended printing buffer buffer systems (for contact printing) • Glycerol up to 40% 150 mM Phosphate, pH 8.5, 5 % • PH stability 5-9 Glycerol, 0,1 mg/ml BSA, 0,01% Sarcosyl Immobilization 8-16 h at 30°C, Over night at 20°C, approximately 65-70% air approximately 65-70% air humidity (saturated sodium humidity (saturated sodium chloride solution) chloride solution) • 20 sek PBST (0.5%) • 3 x 30 min PBST Blocking / Optional Required (to deactivate Deactivation • PBST with 1% BSA, remaining NHS ester groups) 1h, RT PBST (0.01%) with 50 mM • 3 x (briefly) PBST Ethanolamine PBST, 5 min dH2O, 1 min Target Incubation Recommended dilution Recommended dilution buffer buffer for target / anatlyte: for target / anatlyte: • PBST • PBST (0.05%) Washes: Washes: • (Briefly) PBST (0.5%) • PBST (1%), 2 min • 3 x 10 min PBST • PBST (0.5%), 2 min (0.1%) • PBST (0.1%), 2 min • 2 x (briefly) dH2O spin dry (e.g. 1000 RPM, 2 min) Date: 12. Februar 2007