PDE6C Assay Kit

The PDE6C Assay Kit is designed for identification of PDE6C inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE6C to the binding agent.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.The PDE6C inhibitor screening assay kit comes in a convenient 96-well format|with purified PDE6C enzyme|fluorescently labeled PDE6 substrate (cGMP)|binding agent|and PDE assay buffer for 100 enzyme reactions. The key to the PDE6C Assay Kit is the specific binding agent. Using this kit|only two simple steps on a microtiter plate are required for PDE6C reactions. First|the fluorescently labeled cGMP is incubated with PDE6C 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.

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