背景介绍 Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE5A, also known as cGMP-inhibited phosphodiesterase, has been implicated in cardiovascular function and fertility. Phosphodiesterases catalyze the hydrolysis of the phosphodiester bond in dye-labeled cyclic monophosphates. Beads selectively bind the phosphate group in the nucleotide product. This increases the size of the nucleotide relative to unreacted cyclic monophosphate. In the polarization assay, dye molecules with absorption transition vectors parallel to the linearly-polarized excitation light are selectively excited. Dyes attached to the rapidly-rotating cyclic monophosphates will obtain random orientations and emit light with low polarization. Dyes attached to the slowly-rotating nucleotide-bead complexes will not have time to reorient and therefore will emit highly polarized light. 产品介绍 The PDE5A Assay Kit is designed for identification of PDE5A inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE5A to the binding agent. The key to the PDE5A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE5A reactions. First, the fluorescently labeled cGMP is incubated with PDE5A for 1 hour. Second, the binding agent is added to the reaction mix to produce a change in fluorescent polarization that can then be measured using a fluorescence reader equipped for the measurement of fluorescence polarization.