Products from BPS Bioscience require a minimum order value above 400€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. PDE4A is widely expressed in brain tumors and promotes their growth and its inhibitor Rolipram has been identified as a potent anti-inflammatory 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 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 PDE4A Assay Kit is designed for identification of PDE4A inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE4A to the binding agent. The key to the PDE4A1A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE4A1A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE4A1A 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 offluorescence 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 PDE4A1A inhibitor screening assay kit comes in a convenient 96-well format,including purified PDE4A1A enzyme, fluorescently labeled PDE4A substrate (cAMP),binding agent, and PDE assay buffer for 100 enzyme reactions.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: Goldhoff, P., et al., Clin Cancer Res. 2008;14(23):7717-25.Application Reference:
Bioguidage search of active compounds from Waltheria indica L. (Malvaceae) used for asthma and inflammation treatment in Burkina Faso (2014)
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