PDE3A TR-FRET Assay Kit

The PDE3A TR-FRET Assay Kit is designed to provide fast and easy identification of inhibitors of PDE3A (phosphodiesterase 3A) using Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) . The PDE3A TR-FRET Assay Kit comes in a convenient well format, with enough recombinant purified PDE3A enzyme, fluorescently labeled PDE substrate (cAMP), binding agent, and PDE assay buffer for 96 enzyme reactions.Figure 1: Illustration of the assay principle.The reaction uses a fluorescein-conjugated (FAM) cyclic monophosphate nucleotide.  Phosphodiesterase PDE3A catalyzes the hydrolysis of the phosphodiester bond in the cyclic monophosphate nucleotide, releasing the phosphate group for binding. The phosphate group binds to a “Binding Agent” (BA) that is recognized by terbium-labeled donor beads. This results in energy transfer from the terbium to FAM, which emits a fluorescent signal at 520 nm. If unbound to the phosphate group, the terbium-labeled beads emit at λ=490 nm. The fluorescent intensity is measured using a fluorescence plate reader capable of TR-FRET reading and an increase in 520 nm corresponds directly to the activity of PDE3A. In this assay, the intensity of the signal at 520 nm is directly proportional to PDE3A enzymatic activity.Need us to run inhibitor screens or profile your compounds against PDE3A? Check out ourPhosphodiesterase Screening Services.This product has beencited 1 time.