The Bridge-It® PDE Assay is a fluorescence-based, high-throughput screening (HTS) method for measuring cyclic nucleotide phosphodiesterase activity from a purified source and determining the IC 50 of their inhibitors. This assay is based on the format of Bridge-It® cAMP all in one Fluorescence Assay (Mediomics, Cat# 122938).
Adenosine-3’,5’-cyclic monophosphate (cAMP) is an important second messenger which is involved in the modulation of numerous biological processes. The measurement of cAMP is especially important in new drug discovery since cAMP levels are closely related to the activity of one of the major targets for new drug discovery – the G protein-coupled receptors (GPCR). Mediomics fluorescence cAMP assay method for determining the concentrations of cAMP is based on the assay platform design described above. Basically CAP, a bacterial DNA binding protein whose DNA binding activity depends on the presence of cAMP, is used as a highly specific biosensor for measuring cAMP levels. As PDE hydrolyzes cAMP to AMP, the concentration of cAMP decreases, which results in less of the CAP-DNA complex formation and more free “half-site” duplexes with probes (unquenched). The increase in the fluorescence signal is proportional to the PDE activity.
More information at : http://mediomics.com/product/bridge-pde-assay-kit-96-well-format/
*384 well format also available