PDE4D7 TR-FRET BioAssay™ Kit
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 PDE4D7 TR-FRET Assay Kit is designed for identification of inhibitors of PDE4D7 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 PDE4D7 TR-FRET Assay Kit comes in a convenient 96-well format, with purified PDE4D7 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 PDE4D7 activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE4D7 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.
Components
PDE4D7 recombinant enzyme, 1x1ug FAM-Cyclic-3’, 5’-AMP: 20um, 1x50ul PDE assay buffer, 1x25ml Tb donor, 1x30ul Binding Agent, 1x200ul Binding Buffer A, 1x20ml Binding Buffer B, 1x20ml Black, low binding NUNC microtiter plate, 1 plate
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.
