ABSTRACT
Substrate phosphorylation assays are functional assays for kinases that enable the identification of compounds that are competitive inhibitors at the ATP and phosphate-accepting substrate binding sites, as well as allosteric inhibitors. Fluorescence-based detection methods, such as prompt fluorescence, fluorescence polarization (FP), and fluorescence resonance energy transfer (FRET), in conjunction with antibodies specific to the phosphorylated substrates, have become the assays of choice for kinase studies that require high throughput.
