PT  - JOURNAL ARTICLE
AU  - Wooten, Marie W.
TI  - In-Gel Kinase Assay as a Method to Identify Kinase Substrates
AID  - 10.1126/stke.2002.153.pl15
DP  - 2002 Oct 08
TA  - Science's STKE
PG  - pl15--pl15
VI  - 2002
IP  - 153
4099  - http://stke.sciencemag.org/content/2002/153/pl15.short
4100  - http://stke.sciencemag.org/content/2002/153/pl15.full
SO  - Sci. STKE2002 Oct 08; 2002
AB  - 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.