Development of a Transcreener™ kinase assay for protein kinase A and demonstration of concordance of data with a filter-binding assay format

A Transcreenerm kinase fluorescence polarization (FP) assay has been developed for the serine/threonine kinase protein kinase A (PKA). The PKA Transcreenerm kinase assay is an homogenous, competitive antibody-based FP assay that uses Far Red Alexa Fluor 633-labeled adenosine 5' disphosphate (ADP) tracer and mouse monoclonal anti-ADP antibody. The Transcreener (TM) PKA assay was validated with both known PKA inhibitors and library compounds. The TranscreenerTm PKA assay is resistant to low-wavelength (or common) fluorescent interference from small-molecule library compounds and generates IC50 results comparable with current radioactive filter-binding assay.