PDE1C Mouse Assay Kit

Catalog # 61312
Size: 96 reactions

DESCRIPTION: Phosphodiesterases (PDEs) play an important role in dynamic
regulation of cAMP and cGMP signaling. PDE1C is a calmodulin-dependent PDE that is
expressed principally in human myocardium. The PDE1C Assay Kit is designed for
identification of PDE1C inhibitors using fluorescence polarization. The assay is based on
the binding of a fluorescent nucleotide monophosphate generated by PDE1C to the
binding agent.

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 PDE1C (Mouse) Assay Kit comes in a convenient 96-well format, with purified murine
PDE1C enzyme, fluorescently labeled PDE1C substrate (cAMP), binding agent, and PDE
assay buffer for 100 enzyme reactions. The key to the PDE1C (Mouse) Assay Kit is the
specific binding agent. Using this kit, only two simple steps on a microtiter plate are required
for PDE1C reactions. First, the fluorescently labeled cAMP is incubated with a sample
containing PDE1C 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.

COMPONENTS:

MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:
Fluorescent microplate reader capable of measuring fluorescence polarization

APPLICATIONS: Great for studying enzyme kinetics and screening small molecular
inhibitors for drug discovery and HTS applications.

STABILITY: 6 months from date of receipt when stored as directed

REFERENCE(S): Chandrasekaran A, et al., Cell Signal. 2008; 20(1): 139-53.

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