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Nitrocellulose coating
Nexterion® Slide NC
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Protein microarray technology provides a robust way to study protein function in a rapid, economical, and system-wide fashion. Protein microarrays are playing an increasingly important role in proteomic research, as well as moving into the areas of drug discovery and clinical diagnostics. For many of these protein array applications, the slide surface of choice is nitrocellulose. The unique properties of cellulose nitrate polymers, such as high affinity, and high capacity for protein binding, together with their micro-porous structure makes this material an ideal substrate for protein array applications.
To meet the needs of this rapidly expanding market, SCHOTT has joined forces with Sartorius Stedim Biotech, to develop a range of innovative nitrocellulose-coated microarray slides. Sartorius Stedim Biotech is the perfect co-development partner for this product line, as they have been manufacturing cellulose nitrate membranes for over seventy years. Nitrocellulose formulations from Sartorius Stedim Biotech have proven their reliability over the years, in Western Blots, and in diagnostic membranes, such as for pregnancy tests. The diagnostic membrane production expertise of Sartorius Stedim Biotech has been combined with SCHOTT’s microarray slide production know-how to produce protein microarray slides of superior quality, with high reproducibility.
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| Type of
Coating
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Immobilization
method
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Typical
probes
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Recommended application |
Ordering information |
| Nexterion®
product
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Number
of pads
per slide
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Barcode
option
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Item
number
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Slides
per pack
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| Nitrocellulose
3-D "open pore" structure
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Passive absorption through
non-specific interactions
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Proteins Antibody
Glycoproteins
Cell lysate
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Non-contact printer |
Slide NC-N |
1 |
None |
1178057 |
20 |
| Slide NC-N2 |
2 |
None |
1247127 |
5 |
| Slide NC-N3 |
3 |
None |
1247128 |
5 |
| Slide NC-N4 |
4 |
None |
1247129 |
25 |
| Slide NC-N16 |
16 |
None |
1178059 |
5 |
| Nitrocellulose
3-D with “small pore“ structure
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Contact printer
and / or cell lysate arrays
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Slide NC-C |
1 |
None |
1247130 |
20 |
| Slide NC-C2 |
2 |
None |
1247131 |
5 |
| Slide NC-C3 |
3 |
None |
1247132 |
5 |
| Slide NC-C4 |
4 |
None |
1247133 |
25 |
| Slide NC-C16 |
16 |
None |
1247134 |
5 |
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| Key product features |
| • High binding capacity |
| • Excellent spot morphology |
| • Exceptional surface for long-term storage of many printed proteins |
• Performance characteristics of the membranes optimised for use with either contact
• or non-contact microarray printing systems |
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| Typical applications |
| • Sandwich arrays |
| • Antibody profiling |
| • MicroSpot ELISA |
| • Reverse phase assays (NC-C) |
| • Cytokine detection |
| • Protein binder arrays |
| • Protein substrate profiling |
| • Protein activity screening |
| • Protein-DNA
interactions |
| • Biomarker profiling |
| • Serological assays |
| • Enzyme profiling |
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| Suitable probe types |
| • Antibody
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| • Proteins
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| • Glycans
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| • Glycoproteins
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| • Lectins
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| • Antigens |
| • Cell or tissue lysates (NC-C) |
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Structure of Nexterion® nitrocellulose coated slides
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The performance of nitrocellulose coated slides is strongly dependent on the internal membrane structure as well as surface quality. The three-dimensional structure and uniform pore size has been optimized to offer a high binding affinity for proteins, and to better stabilize their active conformation. Proteins are initially attracted to a membrane surface by electrostatic interaction. Long-term attachment is achieved by a combination of hydrogen bonding and hydrophobic interactions. The SCHOTT nitrocellulose slide membranes are manufactured using a newly developed casting process, where solvents are evaporating from a mixture of different cellulose nitrate polymers, and additives, to form the unique micro-porous structure. During manufacture, the thickness, and physical characteristics of the coating are tightly controlled, these together with a surface finishing process allow the reproducible manufacturing of slides with an outstanding consistency and performance. The Nexterion® NC slides use two different types of nitrocellulose formulations developed by Sartorius Stedim Biotech specifically for protein microarray applications.
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There are two main methods used to print microarrays, contact and non-contact. Contact printing systems typically use solid pins, “pin and rings”, slotted pins, split pins / quills, or capillary tubes that must make physical contact with the surface of the membrane to deposit the probes. For the contact method of printing, the nitrocellulose formulation must be robust enough to be able to withstand any potential damage caused by the pins. The internal structure of the membrane must also control the degree of probe “wicking” when the pins touch-down to ensure consistent spot diameters. The NC-C type membranes contain a higher level of “solids”, increasing the binding capacity, and reducing the average size of the micro-pores. This results in a nitrocellulose membrane that has been optimized for contact printing, and in addition, the higher binding capacity offers a superior performance for reverse phase assays that involve printing protein, cell or tissue lysates. Non-contact microarray printing systems, using piezoelectric or syringe-solenoid pressure-assisted deposition methods are often preferred for printing proteins as the deposition volume is very tightly controlled. For non-contact printing method the NC-N type nitrocellulose is the optimal choice.
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Scanning electron microscope photograph of SCHOTT Sartorius Stedim three-dimensional nitrocellulose surface illustrating the uniformity of the two pore structures.
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Product details
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High binding capacity
The three-dimensional micro-porous structure of nitrocellulose provides a
much higher surface area for binding in comparison to two-dimensional
organo-silane coatings. In addition, the nitrocellulose surface binds proteins
in a non-covalent, but irreversible manner and the three-dimensional environment
helps to maintain the proteins in their biologically active conformation.
Robust and smooth membrane surface free of dust and defects
The surface of Nexterion® Slide NC-C is robust enough for use with contact printers, but is sufficiently absorbent to allow a high binding capacity and good spot morphology. The absence of dust and loose particles significantly reduce any risk of pin blockages during contact printing.
Excellent spot morphology
Nitrocellulose is naturally hydrophobic, and this can be a disadvantage in microarraying where it can lead to inconsistent sample volumes being deposited by contact printers. During manufacture the membranes are impregnated with a surfactant, to ensure an optimal surface for microarray printing. The surfactant type and concentration have been carefully selected to ensure a low auto-fluorescence, and to meet the requirements of consistent microspot printing.
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Background signals of Nexterion® Slide NC-N in comparison to a commercially available nitrocellulose coated slide.
Scanned with Axon Scanner Genepix 4000 B, Scan parameters: 532 nm, laser power: 33 %, PMT gain: 500
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Nitrocellulose structure is optimised for the two main types of printing technology
SCHOTT offers two distinct types of nitrocellulose membranes. Nexterion® Slide NC-N for use with non-contact microarray printing systems. Nexterion® Slide NC-C is recommended for use with contact printers. Both types of nitrocellulose membranes are compatible with all detection methods including fluorescent, chemi-luminescent, radiographic, and colorimetric.
Long term stability
Nexterion® NC-C/N slides are ready-to-use from the box, and are stable for over one year under ambient conditions. It is well documented that nitrocellulose provides an excellent surface for the long-term storage of many printed proteins, especially for antibodies and antigens.
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For blocking, hybridization and washing no pre-made Nexterion® reagents are available.
Please check the appropriate slide protocol for recommendations.
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Nexterion® Slide NC-C/N product range
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The conventional format for DNA arrays has become standardized around the microscopic glass slide, and now the same instrumentation is being used to print and scan protein microarrays. The dimensions of the glass slides used for the SCHOTT nitrocellulose slides are industry standard 75.6 x 25.0 x 1.0 mm. The glass foundation is a high quality, extremely flat borosilicate glass with a low intrinsic auto-fluorescence.
SCHOTT offers a choice of either single or multi-pad nitrocellulose coated glass slides with either the NC-C or NC-N nitrocellulose formulations. The spacing and locations of the pads on the standard NC-C/N slides are exactly the same as industry standard nitrocellulose slides allowing the use of conventional slide processing equipment. The propriety method used to create the individual pads eliminates any “edge effect” allowing arrays to be reliably printed very close to the edges. The method is also very flexible, making custom pad formats cost effective.
Single pad slides / Nexterion® Slide NC-C/N
White nitrocellulose-coated glass slides with a single pad 21 by 51 mm.
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Nexterion® Slide NC-C/N2
White nitrocellulose-coated glass slides with 2 pads each 20 by 20 mm.
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Nexterion® Slide NC-C/N3
White nitrocellulose-coated glass slides with 3 pads each 20 by 20 mm. Corner orientation mark.
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Nexterion® Slide NC-C/N4
White nitrocellulose-coated glass slides with 4 pads each 6 by 6 mm. Pad centre-to-centre spacing: 9 mm.
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Nexterion®
Slide NC-C/N16
White nitrocellulose-coated glass slides with 16 pads each 6 by 6 mm. Pad centre-to-centre spacing: 9 mm. Corner orientation mark.
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Accessories
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The use of either single-use gasket chambers, or re-usable silicone incubation
chambers, is recommended for carrying out the post-printing incubation
reactions.
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SCHOTT Nexterion® MPX-4 tray for the rapid processing of up to 4 slides
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Nexterion® MPX-4 tray holder for four slides
The Nexterion® MPX-4 tray (1180496) is designed to hold up to four Nexterion® Slide NC slides. The tray allows the use of automated liquid handling systems, or multi-channel hand-held pipettes. The external dimensions of the MPX-4 tray conform to the SBS microplate standard allowing the high throughput processing of up to 64 arrays simultaneously.
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Dimensions Nexterion® MPX-4 tray
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Compatible accessories
The dimensions and locations of the pads on the SCHOTT single-pad, and multi-pad slides are fully compatible with other commercially available incubation chambers.
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| Item |
SCHOTT product |
Grace Biolabs* |
Whatman** |
Nexterion® Slide NC-C/N
Single pad slide
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SecureSeal* SA2657 Hybridisation chambers.
Internal 53 x 22 mm.
Order code: 622506
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Single-Well Incubation Chamber.
Order code: 10486137
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Nexterion® Slide NC-C/N
2-pad slide
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SecureSeal* SA2X2121 Hybridisation chambers.
Internal 21 x 21 mm.
Order code: 622511
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Dual well incubation chamber
Order code: 10486087
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Nexterion® Slide NC-C/N
3-pad slide
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SecureSeal* SA3S-0.5 Hybridisation chambers.
Internal 20 x 21 mm.
Order code: 622508
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Nexterion® Slide NC-C/N
4 or 16-pad slides
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1. ProPlate* slide module.
Order code: 204862
2. SecureSeal* SA16S-0.5 Hybridisation chambers 16 chambers 7 x 7 mm 9 mm spacing.
Order code: 622510
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16-Well Array Incubation Chamber.
Order code: 10486046
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| Holder for processing single slides |
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Chip Clip** Slide-Holder
Order code: 10486081
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| SBS compliant microplate holder for processing 4 slides |
Nexterion® MPX-4 tray. Reusable.
Order code: 1180496
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Flexwell slide tray
Order code: 204970
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FAST** Frame Multi-Slide. Plate.
Order code: 10486001
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* Trademarks of Grace Biolabs
** Trademarks of Whatman Group |
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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: 1.1
Revision made: New telephone and fax numbers
Revision reason: Update 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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