Immunochemical methods for the rapid screening of the o-glycosidically linked N-acetylglucosamine modification of proteins

Methods Mol Biol. 2008:446:267-80. doi: 10.1007/978-1-60327-084-7_19.

Abstract

for the rapid screening of specific post-translational modifications antibody-based methods are very well suited and applicable without demanding expenditure. Here we describe the immunochemical detection of the O-glycosidically linked cytosolic N-acetylglucosamine modification of proteins, which has attracted increasing interest in the last years. Two different monoclonal antibodies were used in enzyme-linked immunosorbent assays (ELISA), Western blots of 1- and 2- dimension (1D and 2D) separated proteins and immunohistochemical analysis of tissue sections. Slight differences in the recognition of this post-translational epitope by the 2 antibodies are observed and will be discussed.

MeSH terms

  • Acetylglucosamine / chemistry*
  • Animals
  • Blotting, Western
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Glycosides / chemistry*
  • Immunohistochemistry
  • Mice
  • Proteins / chemistry*

Substances

  • Glycosides
  • Proteins
  • Acetylglucosamine