The PDE3B Assay Kit is designed for identification of PDE3B inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE3B to the binding agent. The key to the PDE3B Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE3B reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE3B for 1 hour. Second, 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.
The PDE1C inhibitor screening assay kit comes in a convenient 96-well format, including purified PDE1C enzyme, fluorescently labeled PDE1C substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
Materials Required But Not Supplied
Fluorescent microplate reader capable to measure fluorescence polarization
*The concentration of PDE3B is lot-specific and will be indicated on the tube containing the enzyme.
UniProt #
Q14432
Background
Phosphodiesterases (PDEs) play an important role in dynamic regulation of
cAMP and cGMP signaling. PDE3B, also known as cGMP-inhibited phosphodiesterase, is
involved in mediating the antilipolytic and anti glycogenlytic effects of insulin in adipose and liver
tissues. PDE3B can hydrolyze both cAMP and cGMP.
References
Chandrasekaran, A., et al., Cell Signal. 2008; 20(1):139-53.
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.
Instructions for Use
See assay kit data sheet for detailed protocol.
Applications
Useful for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Shipping Temperature
-80°C (dry ice)
Warnings
This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required partsto read the FP signal. For more information FP technology, visit our Tech Note: FP, assay principles and applications.
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.