In-Gel Activity Assay for Glycosidases.

Authors

Nina V Stourman

Published in

Methods in molecular biology (Clifton, N.J.). Volume 2917. Pages 133-141.

Abstract

Fluorogenic sugar derivatives are useful substrates for the detection of glycosidase activity within a native polyacrylamide gel. When an enzyme cleaves the glycosidic bond, a fluorescent compound is released at the protein's location, forming a band visible under UV light. This method can be used to identify proteins with glycosidase activity within a mixture, evaluate the effect of mutations on enzyme activity, and even provide information about the quaternary structure of a glycosidase. Here, we present a protocol for glycosidase zymography and demonstrate its utility as a tool to probe the active oligomeric form of a novel bacterial β-galactosidase.

PMID:
40347338
Bibliographic data and abstract were imported from PubMed on 11 May 2025.

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