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Three-dimensional thin film coating
Nexterion® Slide P
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SCHOTT specifically developed Nexterion® Slide P as a dedicated slide surface
for printing arrays for comparative genomic hybridization (aCGH), as it is
ideally suited for the covalent immobilization of long amino-modified DNA
molecules such as BAC and PAC clones. Nexterion® Slide P 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. Since its introduction, the slide coating has also been successfully
used for printing antibody and other protein arrays. 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. The robust coating matrix is fully compatible
with commercial microarray printers and scanners. Simple and robust protocols
are available making Nexterion® Slide P 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 |
• Amino-linked BACs, PACs
• Antibodies
• Proteins
• Peptides
• Glycans |
Slide P |
Laser |
1167904 |
25 |
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| Key product features |
| • Ideal substrate for printing amino-modified BACs and antibody microarrays |
| • High probe loading capacity |
| • Exceptionally wide dynamic range |
| • Extremely 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 |
| • Array comparative genomic hybridization (aCGH) |
| • Antibody profiling |
| • Protein expression profiling |
| • Functional protein arrays |
| • Characterization of binding molecules |
| • Biomarker discovery |
| • Substrate profiling |
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| Suitable probe types |
| • Amino-modified BACs, PACs and PCR products |
| • Amino-modified oligonucleotides |
| • Antibodies |
| • Functional proteins such as enzymes or receptors |
| • Peptides |
| • Glycans |
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The coating on the SCHOTT Nexterion® Slide P is a three-dimensional,
hydrophilic polymer 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 P coating has been engineered to exhibit an extremely
low intrinsic non-specific background without the need for blocking. This was
achieved by using a special polymer that is extremely resistant to non-specific
binding. During in-house tests run by SCHOTT, Nexterion® Slide P had the
lowest background signal of any microarray slide coating ever tested. The
polymer coating has a three-dimensional structure; with NHS-ester reactive
groups attached to a hydrophilic polymer backbone. The terminal amino
group of amino-modified nucleic acids react immediately and irreversibly
with the NHS-ester groups to form a covalent bond. Proteins and other complex
bio-molecules bind via by surface-exposed amino-groups. The polymer
coating maintains the immobilized bio-molecules in a quasi-liquid environment
that maintains the protein specificity and chemical conformation. The
three-dimensional polymer structure, combined with the end-point attachment
chemistry, orients the immobilized bio-molecules away from the glass
facilitating the interactions of the attached bio-molecules with their binding
targets in solution.
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Product details
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Highly reproducible coating
Nexterion® Slide P is fabricated using a proprietary thin-film deposition process
developed 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 tightly controlled, class 100 clean
room facility, resulting in coated slides with highly uniform surface properties
and low auto-fluorescence.
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Figure shows scanned images of Nexterion® Slide P
evaluated in an anti-IgG/IgG interaction study
over a range of probe concentrations dissolved in
Nexterion® Spot PB and a phosphate print buffer.
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Packaging and storage
Nexterion® Slide P 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 P is available in packs of 25 slides with code 128 barcodes
enabling automated sample tracking. The identical P coating is also available
in a 16-well slide and 96-well microplate formats. For further information
refer to the section on “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© |
Protein concentration 0.1 to 1 mg/mL |
| Spotting (DNA) |
Nexterion® Spot |
100 mM borate (pH 8.0),
50 % DMSO |
Oligonucleotides: 20 µM |
| Spotting (Peptides) |
- |
10 – 50 % DMSO |
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| Chemical deactivation
of unreacted
NHS-esters |
- |
25 mM ethanolamine in
100 mM
sodium borate buffer (final pH 8.5) |
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| Incubation (Protein) |
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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 |
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Protocol revision
Date: April 2009
Protocol version: 1.1
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: October 2006
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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Contact
SCHOTT Technical Glass
Solutions GmbH
Otto-Schott-Strasse 13 07745 Jena Germany
 | +49 (0)3641/681-4066 |
 | +49 (0)3641/681-4970 |
E-mail
More contacts
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