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SCHOTT North America
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Three-dimensional thin film coating
Nexterion® Slide H
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SCHOTT launched Nexterion® Slide H as a dedicated slide surface for printing
protein microarrays, as it is ideally suited for the covalent immobilization of
peptides and proteins such as antibodies, antibody fragments, enzymes, or
receptors. For many protein microarray applications Nexterion® Slide H has
proven to be a very attractive alternative to the commonly used nitrocellulose
coated slide, especially where low background, or slide transparency are important
considerations. Since its introduction, the slide coating has also been
successfully used with amino-modified oligonucleotides, and has become
the slide of choice for printing amino-linked glycan microarrays.
Carbohydrate
arrays are a rapidly growing area of microarray research and Nexterion®
Slide H is an excellent choice for use in the rapid screening of carbohydrateprotein
interactions. The permeable, polymer coating has a large immobilization
capacity, and helps to preserve the native three-dimensional structure
of complex bio-molecules, thus maintaining conformation and functionality.
Nexterion® Slide H produces excellent signal-to-background ratios and an
exceptionally wide dynamic range compared to conventional “two-dimensional”
coatings through a unique combination of low non-specific binding
characteristics, and high probe loading capacity. Even very low intensity signals,
such as those obtained from low-abundance analytes, or weakly expressed genes can be reliably detected and quantified on Nexterion® Slide H. The
robust coating matrix is fully compatible with commercial microarray printers
and scanners. Simple and robust protocols are available making Nexterion®
Slide H easy-to-use.
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| Type of
coating |
Immobilization
method |
Typical
probes |
Ordering information |
| Nexterion® product |
Barcode
option |
Item
number |
Slides
per pack |
| Thin film 3-D
polymer
surface |
Amine reactive
chemistry
Covalent binding |
• Proteins
• Amino-linked glycans
• Amino-linked oligonucleotides |
Slide H |
Laser |
1070936 |
25 |
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| Key product features |
| • Ideal substrate for printing protein, amino-modified glycans or
oligonucleotide microarrays |
| • High probe loading capacity |
| • Exceptionally wide dynamic range |
| • Very low non-specific binding characteristics |
| • Optimal preservation of native structure and biological activity
of protein probes |
| • Compatible with all common microarray printers and scanners |
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| Typical applications |
| • Proteome expression profiling |
| • Functional protein arrays |
| • Transcriptional profiling |
| • Biomarker discovery |
| • Detection of protein modifications |
| • Carbohydrate–protein interactions |
| • Bacteria serotyping |
| • Immunoassays |
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| Suitable probe types |
| • Antibodies and antibody fragments |
| • Functional proteins such as enzymes or receptors |
| • Small protein fragments such as peptides |
| • Amino-modified oligonucleotides 16 to 70 mers |
| • Amino-linked glycans |
| • Cells |
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The coating on the SCHOTT Nexterion® Slide H consists of a cross-linked,
multi-component polymer layer activated with N-Hydroxysuccinimide (NHS)
esters to provide covalent immobilization of amine groups. All Nexterion®
microarray slides are manufactured from a high quality, low-fluorescence glass
coated with low-fluorescence coatings. However, the non-specific binding of
assay components still remains an important contributor to the off feature
background for many microarray applications. For most types of slide coatings
the post-print processing protocol involves a method of adsorptive blocking
to reduce non-specific binding, however, these procedures are difficult to
perform in a consistent manner.
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The Nexterion® Slide H coating has been engineered to exhibit a very low
intrinsic non-specific background without the need for blocking. This is
achieved by using a polymer based layer that is extremely resistant to nonspecific
binding. The polymer coating has a three-part structure; NHS-ester
reactive groups are attached to the cross-linked hydrophilic polymer layer via
long, flexible spacers. The terminal amino group of amino-modified nucleic
acids and glycans react immediately and irreversibly with the NHS-ester groups
to form a covalent bond. Proteins and other bio-molecules bind via surfaceexposed
amine-groups. The flexible spacers tether the immobilized bio-molecules
in a quasi-liquid environment that maintains the protein specificity and
chemical conformation. The high accessibility of the tethered bio-molecules
facilitates interactions with their binding targets in solution.
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Product details
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Highly reproducible coating
Nexterion® Slide H is fabricated using a proprietary thin-film deposition process
optimized by SCHOTT to produce a uniform and reproducible polymer
coating on one side of a high quality borosilicate glass slide. All slides are
individually examined for physical defects and the presence of particles before
and after coating. The surface is applied in a tightly controlled, class 100 clean
room facility, resulting in coated slides with highly uniform surface properties
and low auto-fluorescence.
Excellent spot morphology and signal-to-background ratios
Nexterion® Slide H provides excellent spot morphologies and reproducible
spot sizes over a wide range of probe concentrations for protein, oligonucleotide
and other bio-molecule microarray applications.
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Figure shows scanned images of Nexterion® Slide H
when evaluated against the market leading threedimensional
competitor slide in an anti-IgG/IgG interaction
study using a range of probe concentrations.
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Figure illustrates the superior signal-to-background
ratios on Nexterion® Slide H when compared to
the market leading three-dimensional slide, and a
conventional two-dimensional coating. This was
demonstrated in two-color hybridization experiments
following the recommended protocols, using
amino-modified oligonucleotide probes.
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Packaging and storage
Nexterion® Slide H are packaged in chemically stable plastic boxes and sealed
under an inert atmosphere. The slides are ready-to-use from the box, and are
stable for 12 months in the sealed packaging when stored at – 20 ºC.
Format
Nexterion® Slide H is available in packs of 25 slides with code 128 barcodes
enabling automated sample tracking. The identical coating is also available
in a 16-well slide format. For further information refer to the section “Multi-well formats”.
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| Protocol step |
Recommended
Nexterion® products |
Alternatives |
Recommended concentrations |
| Spotting (Protein) |
Nexterion® Spot PB |
150 mM phosphate, pH 8.5,
0.01% sarcosyl or Tween20© |
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| Spotting (DNA) |
Nexterion® Spot |
300 mM sodium phosphate (pH 8.5)
containing 0.005 % Tween20® and
0.001 % sarcosyl
150 mM sodium phosphate (pH 8.5)
containing 0.001 % Tween20® |
Oligonucleotides: 20 µM |
| Chemical deactivation
of unreacted
NHS-esters |
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25 mM ethanolamine in
100 mM
sodium borate buffer (final pH 8.5) |
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| Incubation
(Protein) |
- |
137 mM NaCl, 2.7 mM KCl,
4.3 mM Na2HPO4, 1.4 mM KH2PO4,
pH 7.5 with 0.05% Tween20® |
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| Hybridization
(DNA) |
Nexterion® Hyb
(formamide free)
Nexterion® Oligo Hyb
(with formamide) |
2x SSC containing 0.1 % SDS and
0.1 % salmon sperm DNA (formamide
can be added if required) |
Add competitor DNA if appropriate |
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Protocol revision
Date: April 2009
Protocol version: 1.2
Revision made: New telephone and fax numbers
Revision reason: Update contact information
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Protocol revision
Date: April 2009
Protocol version: 1.2
Revision made: New telephone and fax numbers
Revision reason: Update contact information
Date: August 2008
Protocol version: 1.1
Revision made: Changed Tween® concentration (incubation step), pH of blocking solution
Revision reason: Optimization of blocking and incubation
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Please request the MSDS
by completing the form.
(* mandatory fields)
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Contact
Microarray Solutions
SCHOTT North America Inc.
5530 Shepherdsville Road
Louisville, KY 40228
USA
 | +1 -502-657-4411 |
 | +1 -502-966-4976 |
E-mail
More contacts
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