PDE1B Assay Kit

Catalog #: 60311
Size: 96 reactions
$350
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.
Description:
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.
Synonym(s):
inhibitor screening, assay kit, PDE1B
Supplied As:
The PDE1B inhibitor screening assay kit comes in a convenient 96-well format, with purified PDE1B enzyme, fluorescently labeled PDE1B substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
Contraindications:
DMSO >1%, strong acids or bases, ionic detergents, high salt
Format:

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.

Application(s):
Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Reference(s):
Siuciak JA, et al. (2007) Neuropharmacology 53(1):113-24.
Notes:
MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:
Fluorescent microplate reader capable to measure fluorescence polarization
Warning(s):
Avoid freeze/thaw cycles
Scientific Category:
PDE
Product Type:
Assay Kit
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Product Datasheets

60311.pdf