蛋白质芯片 Slide H 应用于探针的制备

3-Dthin film coating 三维薄膜涂层

NEXTERION® Slide H

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.

Overview

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

Immobilization chemistry

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.

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.

Product details

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.

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 and 48-well slide format.