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Curr Protoc Protein Sci. 2001 Nov;Chapter 12:Unit 12.8. doi: 10.1002/0471140864.ps1208s25.

Detection and analysis of proteins modified by O-linked N-acetylglucosamine.

Author information

1
The Johns Hopkins University, Medical School, Baltimore, Maryland, USA.

Abstract

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.

PMID:
18429112
DOI:
10.1002/0471140864.ps1208s25
[Indexed for MEDLINE]

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