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A simple and rapid technique for detecting protein phosphorylation using one‐dimensional isoelectric focusing gels and immunoblot analysis
Author(s) -
Anderson Jeffrey C.,
Peck Scott C.
Publication year - 2008
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2008.03550.x
Subject(s) - isoelectric focusing , phosphorylation , protein phosphorylation , isoelectric point , chromatography , microbiology and biotechnology , biology , biochemistry , chemistry , protein kinase a , enzyme
Summary We report a technique for detecting protein phosphorylation that involves isoelectric focusing in a vertical mini‐gel format followed by immunoblot detection of the target protein. This method uses standard protein gel equipment, allows sensitive detection of protein phosphorylation when phosphospecific antibodies are not available, and provides a stoichiometric measure of phosphorylation. We demonstrate the application of this method for observing phosphorylation of an Arabidopsis thaliana protein in response to biotic stress.