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Aldehydesilane coating
Nexterion® Slide AL
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Nexterion® Slide AL offers a good alternative to Nexterion® Slide E (Epoxysilane
slides) when an efficient covalent, and directed binding of amino-modified
bio-molecules is required. Nexterion® Slide AL is coated with an “active” slide
chemistry coating that chemically binds bio-molecules to provide a high binding
capacity, and minimize non-specific binding. The uniform surface features
aldehyde groups that readily react with primary amines. Both 5’ or 3’ amine
modified PCR products, and oligonucleotides work well with this surface
chemistry. The covalent bond formed at the terminus of the nucleic acid offers
both stability, and maximal base pairing opportunity. The covalent binding
diminishes sample loss during the course of experiments, and permits more
harsh wash steps, which reduce background, and allow for greater sensitivity.
Additional immobilization steps, such as baking or UV cross-linking, are not
required for immobilization. Furthermore, peptides, proteins (such as antibodies),
cells, and tissues can be immobilized on Nexterion® Slide AL via random
binding to amine sites on the probes. The slides are easy to use, and are
compatible with all commercially available arraying and scanning instruments.
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| Type of
coating |
Immobilization
method |
Typical
probes |
Ordering information |
| Nexterion® product |
Barcode
option |
Item
number |
Slides
per pack |
| Aldehydesilane
2-D surface |
Amine reactive chemistry
Covalent binding |
Amino-modified PCR
products, BACs and
oligonucleotides |
Slide AL |
None |
1064874 |
25 |
| Laser |
1064876 |
25 |
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| Key product features |
| • Stable and covalent binding of probes such as amino-modified nucleic
acids, cells, and peptides |
| • Optimal accessibility of probes through specific end-point attachment |
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| Typical applications |
| • Array CGH using whole genome tiling path BAC arrays |
| • miRNA expression profiling |
| • Indel oligonucleotide arrays for genotyping |
| • 16S rRNA-based taxonomic microarray |
| • Protein-DNA interaction studies |
| • Protein-protein interaction studies |
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| Suitable probe types |
| • Amine-PCR products and BACs |
| • Amino-modified oligonucleotides |
| • Small protein fragments such as peptides |
| • Cells and tissues |
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The DNA product is spotted onto the aldehydesilane slide. The primary
amino linkers (NH2) on the DNA attack the aldehyde groups to form covalent
bonds (Schiff’s base). The attachment
is stabilized by a dehydration reaction.
To minimize fluorescent background,
unreacted aldehyde groups are reduced
to non-reactive primary alcohols by
treatment with sodium borohydride.
In addition, this step reduces the double
bond between the probe and surface,
producing an irreversible covalent immobilization.
Additional steps for immobilization,
such as baking, or UV cross-linking,
are not required.
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Product details
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Highly reproducible coating
Nexterion® Slide AL are produced using an innovative process developed by
SCHOTT to produce a consistent and reproducible coating. All slides are individually
examined for physical defects
and the presence of particles before
and after coating. The density of the
aldehyde groups on the coating remain
constant over the entire surface of the
slides, and has been optimized to provide
the optimal binding capacity. The
surface hydrophobicity is tightly controlled
to optimize the performance with
both contact and non-contact printers.
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High intra-slide reproducibility of Nexterion® Slide AL
(20 k array of BSA labelled with Atto 547)
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Packaging and storage
Nexterion® Slide AL are packaged in chemically stable plastic boxes and
sealed in an inert atmosphere. The slides are ready-to-use from the box,
and are stable for 9 months in the sealed packaging when stored at room
temperature.
Format
The slides are available in packs of 25-slides with optional code 128 barcodes
enabling automated sample tracking. The aldehydesilane coating is also available
in 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 |
Additional information |
| Spotting |
Nexterion® Spot |
Nexterion® Spot + detergents
like cetyltrimethylammoniumbromid,
Triton X-100, sarcosyl,
Tween or SDS with a final
concentration between 0.005
and 0.05 % (to increase the
spot size) |
Recommended Spotting
Concentrations:
PCR products: 0.1 – 1 µg/µL
Oligonucleotides: 10 – 20 µM
Peptides: 100 – 500 µg/mL
Recommended Spotting Conditions:
Constant relative humidity
40 – 50% at 20 – 24°C (68 to 75°F).
Spotting solutions may be used with,
or without protein-stabilizing agents |
| 3x SSC |
| 3x SSC + 1.5 M betaine
(low evaporation buffer for
long spotting runs) |
| Chemical
deactivation |
- |
1.0 g NaBH4 in 300 mL PBS
and 100 mL ethanol |
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| Hybridization |
Nexterion® Hyb
(formamide free)
Nexterion® Oligo Hyb
(with formamide) |
3 – 5x SSC + 0.1 % SDS |
Add competitor DNA if appropriate |
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Nexterion® Slide AL Evaluation Kit
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Nexterion® Slide AL is easy to use, but as this type of slide chemistry may be
less familiar to some researchers, a dedicated Nexterion® Slide AL Evaluation
Kit is available. The kit consists of slides with test oligonucleotides and optimized
reagents, allowing new users to carry out a test print and hybridization.
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| Process step |
SCHOTT product |
Quantity |
| Spotting |
Nexterion® Slide AL |
10 slides |
| Nexterion® Spot |
10 mL |
| Hybridization |
Nexterion® Hyb |
10 mL |
| Oligo test probe |
2 nmol |
| Oligo test target |
3 pmol |
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Protocol revision
Date: April 2009
Protocol version: 1.2
Revision made: New telephone and fax numbers
Revision reason: correction contact information
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Protocol revision
Date: April 2009
Protocol version: 1.1
Revision made: New telephone and fax numbers
Revision reason: correction contact information
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Protocol revision
Date: April 2009
Protocol version: 1.2
Revision made: New telephone and fax numbers
Revision reason: correction contact information
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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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