Coimmunoprecipitation assay for the detection of kinase-substrate interactions

Methods Mol Biol. 2003:218:277-84. doi: 10.1385/1-59259-356-9:277.

Abstract

Coimmunoprecipitation is a powerful tool to study protein-protein interactions and can be used to test for the physical association between a known protein kinase and its substrate. In this chapter, the author describes a protocol for the preparation of a cell lysate, the immunoprecipitation of the antigen, and the analysis of the immune complex by gel electrophoresis and immunoblotting with antibodies that recognize the putative associated protein.

MeSH terms

  • Antibodies
  • Cells, Cultured
  • Immunoblotting / methods
  • Precipitin Tests / methods*
  • Protein Kinases / analysis
  • Protein Kinases / immunology
  • Protein Kinases / metabolism*
  • Substrate Specificity

Substances

  • Antibodies
  • Protein Kinases