PDE4D2 TR-FRET Assay 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 PDE4D2 TR-FRET Assay Kit is designed for identification of inhibitors of PDE4D2 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.
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.
Need us to run inhibitor screens or profile your compounds against PDE4D2? Check out our Phosphodiesterase Screening Services.
Fluorescent microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET)
Catalog # | Component | Amount | Storage | |
60048 | PDE4D2 recombinant enzyme | 1 µg | -80°C | Avoid freeze/ thaw cycles! |
60200 | FAM-Cyclic-3′, 5′-AMP: 20 μM | 50 µl | -80°C | |
60393 | PDE assay buffer | 25 ml | -20°C | |
60394 | Tb donor | 50 µl | -80°C | |
60390 | Binding Agent | 200 µl | +4°C | |
78422 | Binding Buffer A | 20 ml | +4°C | |
78423 | Binding Buffer B | 20 ml | +4°C | |
79685 | Black, low ginding microtiter plate | 1 | Room temp. |