Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.14/103739
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- Title
- Small-scale enzymatic digestion of glycoproteins and proteoglycans for analysis of oligosaccharides by LC-MS and FACE Gel Electrophresis
- Related
- Packer, Nicolle H. and Karlsson, Nichlas G.. Glycomics : methods and protocols, p.171-192
- DOI
- 10.1007/978-1-59745-022-5_13
- Related
- Methods in Molecular Biology 534
- Publisher
- USA : Springer; Humana Press
- Date
- 2009
- Author/Creator
- Estrella, Ruby P
- Author/Creator
- Whitelock, John M
- Author/Creator
- Roubin, Rebecca H
- Author/Creator
- Packer, Nicolle H
- Author/Creator
- Karlsson, Niclas G
- Description
- Structural characterization of oligosaccharides from proteoglycans and other glycoproteins is greatly enhanced through the use of mass spectrometry and gel electrophoresis. Sample preparation for these sensitive techniques often requires enzymatic treatments to produce oligosaccharide sequences for subsequent analysis. This chapter describes several small-scale methods for in-gel, on-blot, and in-solution enzymatic digestions in preparation for graphitized carbon liquid chromatography-mass spectrometry (LC-MS) analysis, with specific applications indicated for glycosaminoglycans (GAGs) and N-linked oligosaccharides. In addition, accompanying procedures for oligosaccharide reduction by sodium borohydride, sample desalting via carbon microcolumn, desialylation by sialidase enzyme treatment, and small-scale oligosaccharide species fractionation are included. Fluorophore-assisted carbohydrate electrophoresis (FACE) is another useful method to isolate derivatized oligosaccharides. Overall, the modularity of these techniques provides ease and flexibility for use in conjunction with mass spectrometric and electrophoretic tools for glycomic research studies.
- Description
- 22 page(s)
- Subject Keyword
- glycosaminoglycans
- Subject Keyword
- N-linked oligosaccharides
- Subject Keyword
- fluorophore-assisted carbohydrate electrophoresis
- Subject Keyword
- in-gel enzyme digestion
- Subject Keyword
- on-blot enzyme digestion
- Subject Keyword
- graphitized carbon LC-MS
- Resource Type
- book chapter
- Organisation
- Macquarie University. Dept. of Chemistry and Biomolecular Sciences
- Identifier
- http://hdl.handle.net/1959.14/103739
- Identifier
- ISBN:9781588297747
- Identifier
- ISSN:1064-3745
- Identifier
- mq-rm-2009009140
- Language
- eng
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