MarkerGene™ Carbohydrate Analysis/ Detection Kit
Product ID: M0272
This kit allows the user to quickly estimate and/or compare the composition of carbohydrates obtained from glycoproteins.
Numerous native proteins contain post-transcriptional glycosidic elaboration (sugars) whose structures are dependent both on species and cell type . The characterization of the complex oligosaccharides obtained from these glycoproteins has proven a difficult and time consuming endeavor [2,3]. The Carbohydrate Analysis/Detection Kit is capable of quickly estimating and/or comparing the composition of the carbohydrates in such samples. The Kit provides reagents and protocols for analyzing these carbohydrates through covalent labeling with a fluorescent reagent (1,5-EDANS), and analysis by thin-layer chromatography (TLC). The principle involves enzymatic removal of the oligosaccharides from a native protein (or mixture of reducing sugars), reductive amination of the reducing sugars and analysis of the resultant glycamines using silica-gel two dimensional thin layer chromatography (2D-TLC) or by other well established techniques [2,3]. The advantages of using the 1,5 EDANS fluorophore include its low detection limit, water solubility, pH fluorescence invariance, stability, distinctive fluorescence from protein chromophores, and ability to be detected using normal phase chromatography techniques.
We anticipate the use of this kit to also aid other carbohydrate analyses including HPLC detection and purification, carbohydrate receptor analyses, enzyme inhibition studies, and the like. Additional information on these techniques is available. Custom couplings and determinations can also be arranged if desired. Please contact our technical services department for further information.
- Starr CM, Masada RI, Hague C, Skop E, Klock JC (1996) "Fluorophore-assisted carbohydrate electrophoresis in the separation, analysis, and sequencing of carbohydrates." J. Chrom. A 70(1-2): 295-321.
- Klock JC, Starr CM (1998) "Polyacrylamide Gel Electrophoresis of Fluorophore-Labeled Carbohydrates from Glycoproteins" Meth. Mol. Biol. 76:115-129.
- Feizi T, Galustian C (1999) "Novel oligosaccharide ligands and ligand-processing pathways for the selectins." Trends Biochem Sci. 24:369-372.
- Weitzman S, Scott V, Keegstra K. (1979) "Analysis of glycopeptides as borate complexes by polyacrylamide gel electrophoresis." Anal Biochem. 97(2):438-449.
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