PDE4D7 Assay Kit

The PDE4D7 Assay Kit is a fluorescence polarization (FP), homogeneous, 96-well assay kit designed for the screening and profiling of PDE4D7 (Phosphodiesterase 4D7) inhibitors. This assay takes advantage of a specific fluorescent phosphate-binding nanoparticle. The kit contains enough purified recombinant human PDE4D7, fluorescent probe, PDE assay buffer, Binding Agent, and diluent for 100 reactions. Figure 1: Illustration of the PDE4D7 Assay Kit principle. The assay uses a fluorescein-labeled cyclic adenosine monophosphate (cAMP-FAM for PDE4D7), in which the phosphate group is engaged within the cyclic nucleotide. This is a very small molecule that rotates fast (low FP). PDE4D7 catalyzes the hydrolysis of the phosphodiester bond in the cyclic nucleotide and frees the phosphate group. In a second step the free phosphate group is recognized by a specific phosphate-binding nanobead (Binding Agent) leading to the formation of a large complex, with restricted movement (high FP). FP is proportional to PDE4D7 activity. This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to read the FP signal. For more information FP technology, visit our Tech Note: FP, assay principles and applications. Interested in having us perform the screening and profiling of inhibitors? Visit our Phosphodiesterase Screening & Profiling Services.