Electrophoretic mobility shift assay for the analysis of interactions of jasmonic acid-responsive transcription factors with DNA

Abstract

The electrophoretic mobility shift assay based on nondenaturing polyacrylamide gel electrophoresis is a simple, rapid, and sensitive method for the study of the interaction of transcription factors with DNA in vitro. It relies on a change in the electrophoretic mobility of a DNA fragment when bound to an interacting protein. The assay can be used to test DNA binding of either puri fied or recombinant proteins or uncharacterized binding activities present in crude protein extracts from plant cells or nuclei. It allows the determination of the abundance, af finity, association rate constants, dissociation rate constants, and binding speci ficity of DNA-binding proteins. © Springer Science+Business Media, LLC 2013.