Glycocalyx, literally meaning "sugar coat", is an extracellular polymeric coating surrounding many prokaryotic and eukaryotic cells consisting of glycoproteins, glycolipids, proteoglycans and glycosaminoglycans. The constituents of the glycocalyx play an important role in the process of cell signaling, virus infections, and immunity. However, detection tools for the research of glycobiology are currently in very limited supply.
Raybiotech has pioneered the development of antibody arrays which are now widely applied in the research community with thousands of peer reviewed publications, including Cell and Nature. Taking advantage of advancements in microarray technology developed for antibody arrays, we have developed the first commercially available glycan array for screening protein-carbohydrate interactions. This array will help researchers:
- Identify the glycans binding partners in biological samples
- Identify whether target proteins are carbohydrate binding proteins
- Probe binding of viruses and whole cells to glycans1-3
- Profile the substrate specificity of enzymes (glycosyltransferases, glycosidases, etc.)4-6
- Profile the inflammatory immune response
The 100 synthetic glycans featured in the Glycan Array 100 are the most frequently identified structures showing important binding function in the literature. For example, influenza virus binds to a variety of sialosides with a serotype-specific pattern.7-9 Galectins, which are involved in apoptosis, cell adhesion and T-cell activation suppression, function by binding beta-galactosides.10-12
- Dialysis Vials
- Labeling Reagent
- Stop Solution
- Glycan Array Glass Slide Assembly
- Sample Diluent
- 20X Wash Buffer I
- 20X Wash Buffer II
- Cy3 equivalent dye-conjugated Streptavidin
- Slide Washer/Dryer
- Adhesive device sealer
- Labeling Buffer
- Floating Dialysis Rack
Other Materials Required
- Detection antibodies of interest (For sandwich-based method only)
- Pipettors, pipet tips and other common lab consumables
- Orbital shaker or oscillating rocker
- Aluminum foil
- Distilled or de-ionized water
- 1.5ml Polypropylene microcentrifuge tubes
- Gene microarray scanner or similar laser fluorescence scanner
- Small plastic or glass containers
- KCl, NaCl, KH2PO4 and Na2HPO4 (For label-based method only)
- Beaker, stir plate and stir bar
- Dry the glass slide
- Block array surfaces
- Incubate samples (samples need to be biotinylated for the label-based approach)
- For the the sandwich-based principle, incubate with a detection antibody cocktail
For the label-based principle, incubate the labeled-streptavidin.
- Incubate with Cy3 Equivalent Dye-Streptavidin
- Disassemble the glass slide
- Scan with a gene microarray laser scanner
- Perform densitometry and analysis
Upon receipt, all components of the RayBiotech Glycan Array 100 kit should be stored at -20°C. After initial use, remaining reagents should be stored at 4°C to avoid repeated freeze-thaw cycles and may be stored for up to 3 months (Labeling Reagent, Item B, should be prepared fresh each time before use). Unused glass slides should be kept at -20°C and repeated freeze-thaw cycles should be avoided (slides may be stored for up to 6 months). The entire kit should be used within 6 months of purchase.
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- Paulson, J. C.; de Vries, R. P. Virus Res 2013, 178, 99-113.
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- Xu, R.; de Vries, R. P.; Zhu, X.; Nycholat, C. M.; McBride, R.; Yu, W.; Paulson, J. C.; Wilson, I. A. Science 2013, 342, 1230-1235.
- Xu, R.; McBride, R.; Nycholat, C. M.; Paulson, J. C.; Wilson, I. A. J Virol 2012, 86, 982-990.
- Stevens, J.; Chen, L. M.; Carney, P. J.; Garten, R.; Foust, A.; Le, J.; Pokorny, B. A.; Manojkumar, R.; Silverman, J.; Devis, R.; Rhea, K.; Xu, X.; Bucher, D. J.; Paulson, J. C.; Cox, N. J.; Klimov, A.; Donis, R. O. J Virol 2010, 84, 8287-8299.
- Stowell, S. R.; Arthur, C. M.; Mehta, P.; Slanina, K. A.; Blixt, O.; Leffler, H.; Smith, D. F.; Cummings, R. D. J Biol Chem 2008, 283, 10109-10123.
- Stowell, S. R.; Arthur, C. M.; Slanina, K. A.; Horton, J. R.; Smith, D. F.; Cummings, R. D. J Biol Chem 2008, 283, 20547-20559.
- Carlsson, S.; Oberg, C. T.; Carlsson, M. C.; Sundin, A.; Nilsson, U. J.; Smith, D.; Cummings, R. D.; Almkvist, J.; Karlsson, A.; Leffler, H. Glycobiology 2007, 17, 663-676.