PDE2A Assay Kit
The PDE2A Assay Kit is a fluorescence polarization (FP), homogeneous, 384-well assay designed for the screening and profiling of PDE2A (Phosphodiesterase 2A) inhibitors. This assay takes advantage of a specific fluorescent phosphate-binding nanoparticle. The kit contains enough purified recombinant PDE2A, fluorescent probe, PDE assay buffer, Binding Agent, and diluent for 400 reactions.
Figure 1: Illustration of the PDE2A assay principle.
The assay uses a fluorescein-labeled cyclic adenine monophosphate (cAMP-FAM), in which the phosphate group is engaged within the cyclic nucleotide. This is a very small molecule that rotates fast (low FP). PDE2A catalyzes the hydrolysis of the phosphodiester bond in the cyclic nucleotide and frees the phosphate group. In a second step the free phosphate group is recognized by a specific phosphate-binding nanobead (Binding Agent) leading to the formation of a large complex, with restricted movement (high FP). FP is proportional to PDE activity.
This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) to read the FP signal. For more information FP technology, visit our Tech Note: FP, assay principles and applications.
Need us to run inhibitor screens or profile your compounds against PDE2A? Check out our Phosphodiesterase Screening Services.
Fluorescent plate reader capable of measuring fluorescence polarization.
| Catalog # | Name | Amount | Storage |
| 60020 | PDE2A, GST-Tag * | 1 µg | -80°C |
| 60200 | 20 µM FAM-Cyclic-3′, 5′-AMP | 50 µl | -80°C |
| 60393 | PDE Assay Buffer | 25 ml | -20°C |
| 60390 | Binding Agent | 250 µl | 4°C |
| 60391 | cAMP Binding Agent Diluent | 25 ml | 4°C |
| 79961 | Low binding, black 384-well plate | 1 | Room Temp |
* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.
Phosphodiesterases (PDEs) play an important role in the dynamic regulation of the second messengers cAMP and cGMP signaling, by hydrolyzing them. PDE2A (also known as cGMP-dependent 3',5'-cyclic phosphodiesterase) displays dual specificity for cAMP and cGMP with a bias towards cGMP. It is involved in the regulation of blood pressure and vascular permeability and is a therapeutic target of interest for cardiovascular diseases.
Maurice DH. Front. Biosci. 2005; 10:1221-8.
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