PDE1B Assay Kit

The PDE1B Assay Kit is designed for identification of PDE1B inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE1B to the binding agent. The key to the PDE1B Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE1B reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE1B 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.Need us to run inhibitor screens or profile your compounds against PDE1B? Check out ourPhosphodiesterase Screening Services.

Product Specifications

Background

Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE1B is a calcium-dependent cyclic nucleotide phosphodiesterase that is highly expressed in the striatum. It plays the physiological role in the central nervous system. 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

Q01064

Applications

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

Format

CatalogNumberComponentAmountStorage60011PDE1B recombinant enzyme1 µg-80°CAvoid freeze/thaw cycles!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.