PDE4B2 Assay Kit / BPS Bioquest Product-Datasheet BPS-60343
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PDE4B2 Assay Kit
PDE4B2 Assay Kit Catalog # 60343
Size: 96 reactions DESCRIPTION: Phosphodiesterases (PDEs) play an important role in dynamic regulation ofcAMP and cGMP signaling. PDE4 selective inhibitors are currently in clinical trials for thetreatment of diseases related to inflammatory disorders. Increased expression of PDE4B2was observed in the near-term myometrium. PDE4B2 can be induced by its own substrate,under the control of one of the major utero-contractile agonist, PGE2. The PDE4B2 AssayKit is designed for identification of inhibitors of PDE4B2 using fluorescence polarization. Theassay is based on the binding of a fluorescent nucleotide monophosphate generated byPDE4B2 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 transition vectorsparallel to the linearly-polarized excitation light are selectively excited. Dyes attached to therapidly-rotating cyclic monophosphates will obtain random orientations and emit light withlow polarization. Dyes attached to the slowly-rotating nucleotide-bead complexes will nothave time to reorient and therefore will emit highly polarized light.
The PDE4B2 Assay Kit comes in a convenient 96-well format, with purified PDE4B2enzyme, fluorescently labeled PDE4B2 substrate (cAMP), binding agent, and PDE assaybuffer for 100 enzyme reactions. The key to the PDE4B2 Assay Kit is the specific bindingagent. Using this kit, only two simple steps on a microtiter plate are required for PDE4B2reactions. First, the fluorescently labeled cAMP is incubated with a sample containingPDE4B2 for 1 hour. Second, a binding agent is added to the reaction mix to produce achange in fluorescent polarization that can then be measured using a fluorescence readerequipped for the measurement of fluorescence polarization.
APPLICATIONS: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
APPLICATION REFERENCE:1. Tamam El-Elimat, et al. J. Nat. Prod. 2013, 76:382-387.
REFERENCES: 1. Cullen MD, et al. Bioorg Med Chem Lett. 2008; 18(4):1530-3. 2. Méhats C, et al. BMC Pregnancy Childbirth. 2007; 7 Suppl 1:S12.
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