Immobilized metal ion affinity-based fluorescence polarization (IMAP): advances in kinase screening

The IMAP Fluorescence Polarization technology is a homogeneous antibody-free method for analysis of kinases, phosphatases, and phosphodiesterases. Recent developments to the technology include an enhancement of the reagent system (the Progressive Binding System) that significantly expands the range of useable concentrations of ATP, choices of substrates, and assay configurations. With the new Progressive System, we are able to design multiplexed assays that allow the simultaneous determination of multiple kinase activities. In addition, coupled assays are now possible, allowing the assay of kinases through natural or artificial coupling through kinase cascades.