Download Capillary Electrophoresis of Carbohydrates by Gerald W. Hart (auth.), Pierre Thibault, Susumu Honda (eds.) PDF

By Gerald W. Hart (auth.), Pierre Thibault, Susumu Honda (eds.)

Recent advancements in high-resolution separation strategies 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 suggestions in capillary electrophoresis (CE) for the research of advanced carbohydrates. Written in step by step element to make sure winning experimental effects, those simply reproducible protocols offer tools for pattern guidance 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 sorts of purposes, equivalent to 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 exhibit using CE as an assay strategy for tracking glycosyltransferase job and for deciding upon the organization consistent of carbohydrate-protein interplay. an invaluable appendix describes the constructions of the main mostly encountered carbohydrate residues and oligosaccharides from mammalian and bacterial origins. every one protocol includes unique details on reagents, gear, and invaluable reviews and notes on methods.
well timed and hugely sensible, Capillary Electrophoresis of Carbohydrates offers either amateur and skilled CE analysts with robust instruments for the profitable separation of complicated carbohydrates by way of capillary electrophoresis.

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This amount would normally provide sufficient material for several electrophoretic runs. For isolation of individual oli- 38 Merry and Astrautsova gosaccharides, and carrying out sequencing, additional material is required. 10. A suitable control in which N-glycans have been well characterised should be used as a control for enzyme digestion Examples of such glycoproteins include ribonuclease B, bovine serum fetuin, and haptoglobin (all available from Sigma– Aldrich but there may be some variation in glycosylation profile between different batches).

7. Take care to completely wet each blot strip with the labeling reagent (usually 15 µL are sufficient), cap the tubes, and label the strips for 2–3 h at 65°C in a heating block. 8. Wash the blot strips three times with about 10 mL of 50% acetonitrile, transfer to screw-top Eppendorf tubes, and dry. 9. Add 40 µL (or as much as needed to submerge the strip) of ice-cold 48% aqueous HF acid and dephosphorylate the samples by leaving them for 60–72 h on ice-water. Release of Glycans from Glycoproteins 37 10.

4. , and Kobata, A. (1982) Hydrazinolysis of asparagine-linked sugar chains to produce free oligosaccharides. Methods Enzymol. 83, 263–268. 5. , and Parekh, R. (1993) Use of hydrazine to release in intact and unreduced form both N- and Olinked oligosaccharides from glycoproteins. Biochemistry 32, 679–693. 6. Dwek, R. , Edge, C. , Harvey, D. , Wormald, M. , and Parekh, R. B. (1994) Analysis of glycoprotein-associated oligosaccharides. Annu. Rev. Biochem. 62, 65–100. 7. , Wheeler, S. , Hunter, A.

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