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PDE7A Assay Kit

The PDE7A Assay Kit is designed for identification of PDE7A inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE7A to the binding agent. The key to the PDE7A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE7A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE7A for 1 hour. Second, a binding agent is added to the reaction mix to produce a change in fluorescent polarization. The FP signal is measured using a fluorescent microplate readercapable of measuring fluorescence polarization.Need us to run inhibitor screens or profile your compounds against PDE7A? Check out ourPhosphodiesterase Screening Services.This product has beencited 2 times.

Product Specifications

Background

Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE7A is widely expressed in various tissues including skeletal muscle, T lymphocytes, brain and pancreas and plays and important role in the regulation of osteoblastic differentiation. 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.

UniProt

Q13946-2

Applications

Useful for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.

Format

CatalogNumberComponentAmountStorage60070PDE7A recombinant enzyme1 µg-80°CAvoidfreeze/thawcycles!60200FAM-Cyclic-3′, 5′-AMP: 20 µM50 µl-80°C60393PDE assay buffer25 ml-20°C60390Binding Agent100 µl+4°C60391Binding Agent Diluent (cAMP) 10 ml+4°C79685Black, low binding, microtiter plate1Room Temp.

Shipping Conditions

-80°C

Storage Conditions

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.

Contraindications

DMSO > 1%, strong acids or bases, ionic detergents, high salt

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