PDE1A Assay Kit

Catalog #: 60310
Size: 96 reactions
Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE1A1 is a calmodulin-dependent PDE and expressed in lung and heart. PDE1A1 is believed to regulate vascular smooth muscle contraction and may play a role in sperm function. 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.
The PDE1A Assay Kit is designed for identification of PDE1A inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE1A1 to the binding agent. The key to the PDE1A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE1A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE1A1 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.
Materials Required But Not Provided:
Fluorescent microplate reader capable of measuring fluorescence polarization
inhibitor screening, assay kit, PDE1A
Supplied As:
The PDE1A inhibitor screening assay kit comes in a convenient 96-well format, including purified PDE1A1 enzyme, fluorescently labeled PDE1A substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
DMSO >1%, strong acids or bases, ionic detergents, high salt

Assay Kit Components

Instructions for use:
See assay kit data sheet for detailed protocol.
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.

Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Bender. A., et al. Pharmacol. Rev. 2006; 58: 488-520.
Avoid freeze/thaw cycles
Scientific Category:
Product Type:
Assay Kit
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