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Applications: 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-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.
Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
Description: Phosphodiesterases (PDEs) play an important role in dynamicregulation of cAMP and cGMP signaling. PDE1C is a calmodulin-dependent PDE that isexpressed principally in human myocardium. The key to the PDE1C (Mouse) Assay Kit is thespecific binding agent. Using this kit, only two simple steps on a microtiter plate are requiredfor PDE1C reactions. First, the fluorescently labeled cAMP is incubated with a samplecontaining PDE1C for 1 hour. Second, a binding agent is added to the reaction mix toproduce a change in fluorescent polarization that can then be measured using afluorescence 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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