In-Gel Kinase Assay as a Method to Identify Kinase Substrates

Sci. STKE, 8 October 2002
Vol. 2002, Issue 153, p. pl15
DOI: 10.1126/stke.2002.153.pl15

In-Gel Kinase Assay as a Method to Identify Kinase Substrates

  1. Marie W. Wooten*
  1. Department of Biological Sciences, Program in Cell & Molecular Biosciences, Auburn University, Auburn, AL 36849, USA.
  1. *Contact information. E-mail, wootemw{at}


Phosphorylation of proteins by kinases is central to many cellular processes, including signal transduction. Thus, assays to identify or characterize kinases are a key tool for research in this area. Kinase substrates can be incorporated into polyacrylamide gels and used to characterize kinase activity in mixed samples. This methodology can be adapted for the identification of novel kinase-substrates or kinase-kinases that participate in the regulation of cell signaling. Here, I review the rationale and principles of an in-gel kinase assay. This strategy relies on co-polymerization of a substrate within the gel matrix, followed by detection of enzymatic activity in situ. The following Protocol provides a detailed method for performing the in-gel kinase assay and discusses the uses of the assay to evaluate kinase activity in the context of proliferation, differentiation, and survival pathways.


M. W. Wooten, In-Gel Kinase Assay as a Method to Identify Kinase Substrates. Sci. STKE 2002, pl15 (2002).

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