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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 dynamicregulation of cAMP and cGMP signaling. PDE10A is a dual substrate PDE highlyexpressed in striatal medium spiny neurons. PDE10A inhibitors can improve thecognitive symptoms of schizophrenia, and exhibit potential therapeutic value forHuntington's Disease. 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: The Mouse PDE7A Assay Kit is designed for identification of inhibitors of Mouse PDE7A using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by Mouse PDE7A to the binding agent.The key to the Mouse PDE7A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for Mouse PDE7A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing Mouse PDE7A 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 Mouse PDE7A Assay Kit comes in a convenient 96-well format, with purified Mouse PDE7A enzyme, fluorescently labeled substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Yang, G., McIntyre, K.W., Townsend, R.M.,et al., Phosphodiesterase 7A-Deficient Mice Have Functional T Cells, J. Immunology December 15, 2003, 171 (12):6414-6420.2. Lee, R., Wolda, S., Moon, E., Esselstyn, J., Hertel, C., Lerner, A. PDE7A is expressed in human B-lymphocytes and is up-regulated by elevation of intracellular cAMP, Cellular Signalling, 2002, 14(3): 277-284.
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