Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE10A is a dual substrate PDE highly expressed in striatal medium spiny neurons. PDE10A inhibitors can improve the cognitive symptoms of schizophrenia, and exhibit potential therapeutic value for Huntington’s disease. PDE10A2 is located in cytosol, whereas PDE10A2 is a membrane-associated protein.
The PDE10A2 TR-FRET BioAssay™ Kit is designed for identification of inhibitors of PDE10A2 using TR-FRET (Time Resolved Fluorescence Resonance Energy Transfer) technology. The assay is based on the generation of FAM-labeled nucleotide monophosphates by the phosphodiesterase. These phosphate groups bind to terbium-labeled nanoparticles, resulting in energy transfer from the terbium to the FAM, which emits a fluorescent signal at 520 nm. The change in fluorescent intensity can be easily measured using a fluorescence plate reader. The PDE10A2 TR-FRET Assay Kit comes in a convenient 96-well format, with purified PDE10A2 enzyme, fluorescently labeled PDE substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions. Using this kit, only two simple steps on a microtiter plate are required for the PDE10A2 activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE10A2 for 1 hour. Second, a binding agent and a terbium donor are added to the reaction mix and incubated for 1 hour. Then, fluorescence intensity can be measured using a fluorescence reader.
Applications:Great for screening small molecular inhibitors for drug discovery and HTS applications
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
