Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4248

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Monosaccharide Analysis of Phosphorylated Sugars by Methanolysis and Diazomethane Methylation

David Oxley, Graeme Currie, and Antony Bacic

This protocol was adapted from "Analysis of Carbohydrate from Glycoproteins," contributed by David Oxley, Graeme Currie, and Antony Bacic, Chapter 25, in Purifying Proteins for Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2004.

The first 100 words of the full text of this article appear below.


INTRODUCTION

This protocol presents a method that allows rapid determination of the monosaccharide composition of glycans, glycoproteins, and proteoglycans that contain (or are suspected to contain) phosphorylated sugars. (For samples that do not contain phosphorylated sugars see Monosaccharide Analysis by Methanolysis.)


MATERIALS

Reagents

caution Acetic anhydride

cautiont-Butyl alcohol (redistilled)

caution Diazomethane

Prepare diazomethane according to the manufacturer’s instructions, with a specialized apparatus such as one available from Aldrich Chemical Company. The method is outlined in Org. Synth. Coll., Vol. 4, 1963, p. 250. (http://www.orgsyn.org).

WARNING:

Diazomethane is toxic and an extremely sensitive explosive when dry. Handle with extreme care. Use . . . [Full Text of this Article]

Equipment


METHOD


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Related Protocol

Monosaccharide Analysis by Methanolysis
David Oxley, Graeme Currie, and Antony Bacic
CSH Protocols 2006: 4247. [Extract] [Full Text]