Chapter title |
Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis
|
---|---|
Chapter number | 17 |
Book title |
Protein-Carbohydrate Interactions
|
Published in |
Methods in molecular biology, April 2017
|
DOI | 10.1007/978-1-4939-6899-2_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6898-5, 978-1-4939-6899-2
|
Authors |
Robb, Mélissa, Hobbs, Joanne K., Boraston, Alisdair B., Mélissa Robb, Joanne K. Hobbs, Alisdair B. Boraston |
Editors |
D. Wade Abbott, Alicia Lammerts van Bueren |
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. |
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