PDE1B TR-FRET Assay Kit
Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE1B is a calcium/calmodulin-dependent cyclic nucleotide phosphodiesterase that is highly expressed in the striatum. It plays a physiological role in the central nervous system, and PDE1B activity has been linked to impaired cognition and spatial learning.
The PDE1B TR-FRET Assay Kit is designed for identification of inhibitors of PDE1B using TR-FRET (Time Resolved Fluorescence Resonance Energy Transfer) technology. The assay is based on the generation of FAM-labeled nucleotide monophosphates by the PDE1B. 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 PDE1B activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE1B for 1 hour. Second, a binding agent and a terbium donor are added to the reaction mix and incubated for 30 minutes. Then, fluorescence intensity can be measured using a fluorescence reader.
Need us to run inhibitor screens or profile your compounds against PDE1B? Check out our Phosphodiesterase Screening Services.
Catalog # | Component | Amount | Storage | |
60011 | PDE1B recombinant enzyme | 1 µg | -80°C | Avoid freeze/ thaw cycles! |
60200 | FAM-Cyclic-3′, 5′-AMP: 20 µM | 20 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 binding microtiter plate | 1 | Room Temp |