Introduction
Recent technological advances by RayBiotech have enabled the largest commercially available glycosylation antibody array to date. With the Human Glycosylation Array 8000, researchers can now efficiently obtain glycosylation profiles in their samples. The glycosylation profiles of 8000 human proteins can be simultaneously detected, including, but not limited to, cytokines, chemokines, adipokines, growth factors, angiogenic factors, proteases, soluble receptors, soluble adhesion molecules in cell culture supernatants, serum, plasma and other samples types.
Capture antibodies are printed onto glass slides and the glycans on these capture antibodies are removed. The glass slide arrays come pre-blocked and are ready to be incubated with samples. After incubation with samples and washing to remove unbound proteins, five unique biotin-labeled lectins are incubated with the array to allow binding to the glycans from captured target proteins. Streptavidin-conjugated fluorescent dye (Cy3 equivalent) is then added to the array. Finally, the glass slide is dried and laser fluorescence scanning is used to visualize the signals. These signals are then compared to the array map to identify glycosylated proteins present in the samples.
Product Features
- High density screening
- High sensitivity and specificity
- Low sample volume (20-100 µl per array)
- Large dynamic range of detection
- Compatible with most sample types
- Accurate and reproducible
Target Names
Biotin-Labeled Lectin Mixture |
| | Lectin Name | Sugar specificity |
1 | Concanavalin A | αMan, αGlc |
2 | Dolichos Biflorus Agglutinin | αGalNAc |
3 | Peanut Agglutinin | Galβ3GalNAc |
4 | Ulex Europaeus Lectin 1 | αFuc |
5 | Wheat Germ Agglutinin | GlcNAc |
Application Data/Notes
Typical Results
The following figure shows the RayBio
® Human Glycosylation Antibody Array 507 probed with conditioned media sample. The images were captured using a Axon GenePix laser scanner. The strong signals in row 20 and the upper left and lower right corners of each array are Positive Controls, which can be used to identify the orientation and help normalize the results between arrays.
Suggested Applications
- Identify and profile the glycans in their samples
- Determine whether their biomarker of interest has glycan moieties
- Determine relative expression levels
Kit Components
- Human Glycosylation Array Glass Slides
- Sample Diluent
- 20X Wash Buffer I
- 20X Wash Buffer II
- Cy3 equivalent dye-conjugated Streptavidin
- Biotin-labeled Lectin Mixture
- Adhesive device sealer
- Manual
Protocol Outline
- Dry the glass slide
- Incubate samples
- Incubate with biotin lectin mixture
- Incubate with Cy3 Equivalent Dye-Streptavidin
- Disassemble the glass slide
- Scan with a gene microarray laser scanner
- Perform densitometry and analysis
Storage/Stability
Upon receipt, all components of the Human Glycosylation Array 1000 kit should be stored at -20°C. Once thawed, the glass slide and Cy3 equivalent dye-conjugated Streptavidin should be kept at -20°C and all other components may be stored at 4°C. The entire kit should be used within 6 months of purchase.
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