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Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Background: 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. PDE10A1 is located in cytosol, whereas PDE10A2 is a membrane-associated protein. 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.
Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
Description: The PDE10A2 TR-FRET Assay Kit is designed for identification of inhibitors of PDE10A2 using TR-FRET (Time Resolved Fluorescence Resonance Energy Transfer) technology. 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.
Storage Stability: At least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Supplied As: 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.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Akinori Nishi and Gretchen L. Snyder. J Pharmacol Sci., 2010, 114: 6 - 16.
2. Kenji Omori and Jun Kotera. Circulation Research. 2007; 100: 309-327.
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