Background:Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE4D is a regulator of airway smooth-muscle contractility, and has been identified as a potential risk predictor for ischemic stroke. Additionally, PDE4D has been associated with asthma pathophysiology and bone formation. The PDE4D gene encodes at least 9 different isoforms. 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. Description:The PDE4D2 TR-FRET Assay Kit is designed for identification of inhibitors of PDE4D2 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 PDE4D2 activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE4D2 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. Synonym(s): inhibitor screening, FRET assay kit, PDE4D2 Supplied As: The PDE4D2 TR-FRET Assay Kit comes in a convenient 96-well format, with purified PDE4D2 enzyme, fluorescently labeled PDE substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions. Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt Format: Instructions for use: See assay kit data sheet for detailed protocol. 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.

Application(s): Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications. Reference(s): Bender. A., et al. Pharmacol. Rev. 2006; 58: 488-520. Warning(s): Avoid freeze/thaw cycles Scientific Category: PDE Product Type: Assay Kit Data shown is lot-specific. Contact us for specific information on other lots.

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