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Application: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Background: Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE1C is a calmodulin-dependent PDE that is expressed principally in myocardium. Phosphodiesterases catalyze the hydrolysis of the phosphodiester bond in dye-labeled cyclic monophosphates. Beads selectively bind the phosphate group in the nucleotide product. This increases the size of the nucleotide relative to unreacted cyclic monophosphate. In the polarization assay, dye molecules with absorption transition vectors parallel to the linearly-polarized excitation light are selectively excited. Dyes attached to the rapidly-rotating cyclic monophosphates will obtain random orientations and emit light with low polarization. Dyes attached to the slowly-rotating nucleotide-bead complexes will not have time to reorient and therefore will emit highly polarized light.
Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
Description: Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE1C is a calmodulin-dependent PDE that is expressed principally in human myocardium. The key to the PDE1C (Mouse) Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE1C reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE1C for 1 hour. Second, a binding agent is added to the reaction mix to produce a change in fluorescent polarization that can then be measured using a fluorescence reader equipped for the measurement of fluorescence polarization.
Storage Stability: At least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Supplied As: The PDE1C (Mouse) Assay Kit comes in a convenient 96-well format, with purified murinePDE1C enzyme, fluorescently labeled PDE1C substrate (cAMP), binding agent, and PDEassay buffer for 100 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Chandrasekaran A, et al., Cell Signal. 2008; 20(1): 139-53.
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