Comparison of luminescence ADP production assay and radiometric scintillation proximity assay for Cdc7 kinase.

ABSTRACT: Several assay technologies have been successfully adapted and used in HTS to screen for protein kinase inhibitors; however, emerging comparative analysis studies report very low hit overlap between the different technologies, which challenges the working assumption that hit identification is not dependent on the assay method of choice. To help address this issue, we performed two screens on the cancer target, Cdc7-Dbf4 heterodimeric protein kinase, using a direct assay detection method measuring [(33)P]-phosphate incorporation into the substrate and an indirect method measuring residual ADP production using luminescence. We conducted the two screens under similar conditions, where in one, we measured [(33)P]-phosphate incorporation using scintillation proximity assay (SPA), and in the othe

SUBMITTER: Takagi T 

PROVIDER: S-EPMC3643815 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature