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Updated: Oct 4, 2025

Detection of Protein Palmitoylation in Cultured Hippocampal Neurons by Immunoprecipitation and Acyl-Biotin Exchange ABE
Published on: February 18, 2013
Immunoprecipitation and Western blot-based detection of protein O-GlcNAcylation in cells
Oumaima Ahmed1, Malik Affar1, Louis Masclef1
1Maisonneuve-Rosemont Hospital Research Center and Department of Medicine, University of Montréal, Montréal, QC H3C 3J7, Canada.
Abstract:
Detection of protein O-GlcNAcylation could be challenging. By using the host-cell factor 1 (HCF-1), a known O-GlcNAcylated protein, we immunoprecipitated HCF-1 from transfected HEK293T cells or endogenous HCF-1 from HeLa cells to detect its O-GlcNAc levels by Western blotting. We also take advantage of RNAi or chemical inhibitors to modulate OGT and OGA activities before HCF-1 immunoprecipitation. For complete details on the use and execution of this protocol, please refer to Daou et al. (2011).
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