PDE2A assay kit / BPS Bioscience Product-Datasheet BPS-60320

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PDE2A assay kit

PDE2A Assay KitCatalog # 60320Size: 96 reactions

DESCRIPTION: Phosphodiesterases (PDEs) play an important role in dynamicregulation of cAMP and cGMP signaling. PDE2A, also known as cGMP-stimulatedphosphodiesterase, hydrolyzes cyclic nucleotides cAMP (Km = 2.4 μM) and cGMP, and isinvolved in the regulation of blood pressure and fluid homeostasis. The PDE2A Assay Kit isdesigned for identification of PDE2A1 inhibitors using fluorescence polarization. Theassay is based on the binding of a fluorescent nucleotide monophosphate generated byPDE2A1 to the binding agent.

Phosphodiesterases catalyze the hydrolysis of the phosphodiester bond in dye-labeledcyclic monophosphates. Beads selectively bind the phosphate group in the nucleotideproduct. This increases the size of the nucleotide relative to unreacted cyclicmonophosphate. In the polarization assay, dye molecules with absorption transitionvectors parallel to the linearly-polarized excitation light are selectively excited. Dyesattached to the rapidly-rotating cyclic monophosphates will obtain random orientationsand emit light with low polarization. Dyes attached to the slowly-rotating nucleotide-beadcomplexes will not have time to reorient and therefore will emit highly polarized light.

The PDE2A inhibitor screening assay kit comes in a convenient 96-well format, withpurified PDE2A1 enzyme, fluorescently labeled PDE2A1 substrate (cAMP), binding agent,and PDE assay buffer for 100 enzyme reactions. The key to the PDE2A Assay Kit is thespecific binding agent. Using this kit, only two simple steps on a microtiter plate arerequired for PDE2A1 reactions. First, the fluorescently labeled cAMP is incubated with asample containing PDE2A1 for 1 hour. Second, binding agent is added to the reaction mix toproduce a change in fluorescent polarization that can then be measured using afluorescence readerequipped for the measurement of fluorescence polarization.

COMPONENTS: PDE2A assay kit components

APPLICATIONS: Great for studying enzyme kinetics and screening small molecularinhibitors for drug discovery and HTS applications.

REFERENCE: Maurice DH. Front. Biosci. 2005; 10:1221-8.

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