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

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


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.

MeSH terms

  • Acetylglucosamine / chemistry*
  • Chromatography, Affinity
  • Immunoblotting
  • Lectins / chemistry
  • Protein Processing, Post-Translational
  • Proteins / analysis*
  • Proteins / chemistry*
  • Proteins / isolation & purification


  • Lectins
  • Proteins
  • Acetylglucosamine