Coimmunoprecipitation assay for the detection of kinase-substrate interactions.
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.
Comai
Lucio
L
Department of Molecular Microbiology and Immunology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
D000906:Antibodies; D002478:Cells, Cultured; D015151:Immunoblotting / Q000379:methods; D011233:Precipitin Tests / Q000379:methods*; D011494:Protein Kinases / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D013379:Substrate Specificity
D000906:Antibodies; D011494:Protein Kinases