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Detailed Structural Analysis of N-Glycans Released From Glycoproteins in SDS-PAGE Gel Bands Using HPLC Combined With Exoglycosidase Array Digestions

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 347))

Abstract

In contrast to the linear sequences of protein and DNA, oligosaccharides are branched structures. In addition, almost all glycoproteins consist of a heterogeneous collection of differently glycosylated variants. Glycan analysis therefore requires high-resolution separation techniques that can provide detailed structural analysis, including both monosaccharide sequence and linkage information. This chapter describes how a combination of high-performance liquid chromatography (HPLC) and exoglycosidase enzyme array digestions can deliver quantitative glycan analysis of sugars released from glycoproteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel bands by matching HPLC elution positions with a database of standard glycans.

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© 2006 Humana Press Inc.

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Royle, L., Radcliffe, C.M., Dwek, R.A., Rudd, P.M. (2006). Detailed Structural Analysis of N-Glycans Released From Glycoproteins in SDS-PAGE Gel Bands Using HPLC Combined With Exoglycosidase Array Digestions. In: Brockhausen, I. (eds) Glycobiology Protocols. Methods in Molecular Biology, vol 347. Humana Press. https://doi.org/10.1385/1-59745-167-3:125

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  • DOI: https://doi.org/10.1385/1-59745-167-3:125

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-553-8

  • Online ISBN: 978-1-59745-167-3

  • eBook Packages: Springer Protocols

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