UNIT 12.8 Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine
Published Online: 1 NOV 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Zachara, N. E., Cole, R. N., Hart, G. W. and Gao, Y. 2001. Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine. Current Protocols in Protein Science. 25:12.8:12.8.1–12.8.25.
- Published Online: 1 NOV 2001
- Published Print: SEP 2001
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First, a protocol for increasing the stoichiometry of O-GlcNAc on proteins is given. This is followed by simple techniques for the detection/screening of O-GlcNAc-modified proteins either by immunoblotting or lectin affinity chromatography. Separate protocols verify that the glycan is O-linked GlcNAc. These methods are followed by protocols for more comprehensive analysis of O-GlcNAc modified proteins, including labeling of O-GlcNAc residues with [3H]Gal, and subsequent product analysis. The final two protocols assay for O-GlcNAc transferase and O-GlcNAcase activity, respectively.