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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. PDE7B is highly expressed in the pancreasand in other various tissues. Inhibition of PDE7B leads to induction of apoptosis. 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 PDE7B Assay Kit is designed for identification of inhibitors of Mouse PDE7B using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by Mouse PDE7B to the binding agent. The key to the Mouse PDE7B Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for Mouse PDE7B reactions. First, the fluorescently labeled cAMP is incubated with a sample containing Mouse PDE7B 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 PDE7B Assay Kit comes in a convenient 96-well format, with purified Mouse PDE7B 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. Brooks, M.D., Jackson, E., et al. PDE7B Is a Novel, Prognostically Significant Mediator of Glioblastoma Growth Whose Expression Is Regulated by Endothelial Cells. PLOS ONE. 2014 9(9): e107397.2. Gardner, C., Robas, N., et al. Cloning and Characterization of the Human and Mouse PDE7B, a Novel cAMP-Specific Cyclic Nucleotide Phosphodiesterase, Biochemical and Biophysical Research Communications, 2000, 272(1): 186-192.
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