Our Glycan Sequencing Kit includes the enzymes and buffer required to sequence ten isolated, N-linked oligosaccharides.
Neuraminidase from Arthrobacter ureafaciens – 80 µl (E-S001)
Beta-Galactosidase from Streptococcus pneumoniae – 60 µl (E-BG07)
N-Acetylglucosaminidase from Streptococcus pneumoniae) – 40 µl (E-GL01)
Alpha-Mannosidase from Jack Bean – 20 µl (E-AM01)
Core Alpha-Mannosidase from X. manihotis) – 10 µl (E-AM02)
5X Reaction buffer – 400 µl
Many methods of analysis are available, including HPLC, gel electrophoresis, HPAEC, capillary electrophoresis, and mass spectrometry. For more information on these methods, please contact us.
Storage Store kit at 4°C. Do not freeze.
The Glycan Sequencing Kit is stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.
Purity All QA-Bio Enzymes are tested for contaminating protease by incubating 10 μg of denatured BSA with 2 μl of enzyme at 37°C for 24 hours. SDS-PAGE analysis of the treated BSA shows no evidence of degradation.
The production host strains for our recombinant enzymes have been extensively tested and do not produce any detectable glycosidases. Enzymes purified from native sources are tested for contaminating exoglycosidases The absence of exoglycosidase contaminants is confirmed by extended incubations with the corresponding pNP-glycosides.
Glycan Sequencing Kit References
Dwek, R. A., C. J. Edge, D. J. Harvey, M. R. Wormald and R. B. Parekh. Analysis of glycoprotein-associated oligosaccharides. Ann Rev Biochem 62: 65-100 (1993).
Kobata, A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem 100: 1-14 (1979).
O’Neil Enzymatic Release of Oligosaccharides from Glycoproteins for Chromatographic and Electrophoretic Analysis. J Chromatogr A . . . 720:1-2, 201-15(1996).
Rudd et al. Oligosaccharide Sequencing Technology. Nature. 388: 6638, 205-7(1997).
Schaumann et al. Analytical Technique for Studying the Structure Of Glycoprotein N-Glycans. J. of Chromatography. 646: 227- 234(1993).
Jackson. The Analysis of Fluorophore-Labeled Carbohydrates by Polyacrylamide Gel Electrophoresis. Mol. Biotechnol. 5:2 101- 23(1996).
Prime, S., et al. Oligosaccharide sequencing based on exo- and endoglycosidase digestion and liquid chromatographic analysis of the products. J Chromatogr A. 720: 263-274 (1996).
Anumula, K.R. & Du, P. Characterization of Carbohydrates using Highly Fluorescent 2- Aminobenzoic acid Tag Following Gel Electrophoresis of Glycoproteins. Anal. Biochem. 15:236- 242(1999).
Geyer Het al. Structural analysis of glycoconjugates by on-target enzymatic digestion and MALDI-TOF-MS. Anal Chem. 71(2):476-82(1999).