Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis

Mélissa Robb, Joanne K Hobbs, Alisdair B Boraston

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

Fluorophore-assisted carbohydrate electrophoresis (FACE) is a method in which a fluorophore is covalently attached to the reducing end of carbohydrates, thereby allowing visualization following high-resolution separation by electrophoresis. This method can be used for carbohydrate profiling and sequencing, as well as for the determination of the specificity of carbohydrate-active enzymes. Here, we describe and demonstrate the use of FACE to separate and visualize the glycans released following digestion of oligosaccharides by glycoside hydrolases (GHs) using two examples: (1) the digestion of chitobiose by the streptococcal β-hexosaminidase GH20C, and (2) the digestion of glycogen by the GH13 member SpuA.

Original languageEnglish
Pages (from-to)215-221
Number of pages7
JournalMethods in Molecular Biology
Volume1588
DOIs
Publication statusPublished - 2017

Keywords

  • Carbohydrates/analysis
  • Disaccharides/analysis
  • Electrophoresis/methods
  • Glycogen/analysis
  • Glycoside Hydrolases/metabolism
  • Pyrenes/chemistry

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