By Gerald W. Hart (auth.), Pierre Thibault, Susumu Honda (eds.)
Recent advancements in high-resolution separation innovations in accordance with capillary-scale chromatography and electrophoresis have reworked the research of unfastened and conjugated mono- and oligosaccharides. In Capillary Electrophoresis of Carbohydrates, hands-on specialists describe state of the art ideas in capillary electrophoresis (CE) for the research of complicated carbohydrates. Written in step by step element to make sure profitable experimental effects, those simply reproducible protocols supply tools for pattern practise and research of mono- and oligosaccharides, glycoproteins, and glycoconjugates. diversified CE separation codecs, together with capillary quarter electrophoresis (CZE), capillary isoelectric focusing (CIEF), capillary isotachophoresis (CITP), capillary gel electrophoresis (CGE), and micellar electrokinetic chromatography (MEKC), are provided for various forms of purposes, resembling the research of glycoforms, the separation of enantiomers, and the answer of oligosaccharides. The separation of local derivatized carbohydrates is gifted utilizing UV, fluorescence, and mass spectrometric detection. different functions reveal using CE as an assay technique for tracking glycosyltransferase task and for settling on the organization consistent of carbohydrate-protein interplay. an invaluable appendix describes the constructions of the main ordinarily encountered carbohydrate residues and oligosaccharides from mammalian and bacterial origins. each one protocol includes special info on reagents, gear, and precious reviews and notes on methods.
well timed and hugely useful, Capillary Electrophoresis of Carbohydrates offers either amateur and skilled CE analysts with robust instruments for the profitable separation of complicated carbohydrates through capillary electrophoresis.
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Extra resources for Capillary Electrophoresis of Carbohydrates
05% sodium azide. 8. It is recommended that at least 250 µg of glycoprotein is used for analysis. 9. The actual amount of glycoprotein required for profiling will depend on the size of the protein, the amount of glycosylation, and the degree of oligosaccharide heterogeneity. In general, the amount of glycoprotein required increases with the size of the protein or the degree of heterogeneity and decreases with the percent of glycosylation. As a general guideline, one would start with approx 50–100 µg to profile the N-linked oligosaccharides of a 60-kDa glycoprotein that contains 10–20% carbohydrate by weight.
10. Tarentino, A. , Schrader, W. , Changchien, L. , and Plummer, T. , Jr. (1992) Multiple endoglycosidase (Endo) F activities expressed by Flavobacterium meningosepticum. Endo F1: molecular cloning, primary sequence, and structural relationship to Endo H. J. Biol. Chem. 267, 3868–3872. 11. Plummer, T. , Jr. and Tarentino, A. L. (1991) Purification of the oligosaccharide-cleaving enzymes of Flavobacterium meningosepticum. Glycobiology 1, 257–263. 12. Trimble, R. B. and Tarentino, A. L. (1991) Identification of distinct endoglycosidase (endo) activities in Flavobacterium meningosepticum: endo F1, endo F2, and endo F3.
Introduction The monosaccharides comprise a number of isomers and homologues having very similar structures, basically composed of hydroxymethyl or hydroxymethine groups linked to each other. In reducing monosaccharides the hemiacetal group is at the reducing ends in addition to these polyhydroxylated basal structures. There are other series of monosaccharides with the ketal or carboxyl group instead of or in addition to the hemiacetal group. Substitution of the hydroxyl group by the hydrogen atom or the amino group produces the deoxy sugar or the amino sugar, respectively.