PDE11A Assay Kit

Catalog # 60411
Size: 96 reactions

DESCRIPTION: Phosphodiesterases (PDEs) are involved in the dynamic regulation of
cAMP and cGMP signaling. PDE11A has been observed in skeletal muscle, prostate,
testis, salivary gland, thyroid gland, and liver. PDE11A may play role in the CNS with an
association to major depressive disorder. The PDE11A Assay Kit is designed for
identification of PDE11A inhibitors using fluorescence polarization. The assay is based
on the binding of a fluorescent nucleotide monophosphate generated by PDE11A 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 PDE11A inhibitor screening assay kit comes in a convenient 96-well format,
including purified PDE11A enzyme, fluorescently labeled PDE11A substrate (cAMP),
binding agent, and PDE assay buffer for 100 enzyme reactions. The key to the PDE11A
Assay Kit is the specific binding agent. Using this kit, only two simple steps on a
microtiter plate are required for PDE11A reactions. First, the fluorescently labeled cAMP
is incubated with a sample containing PDE11A 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.

COMPONENTS:

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): Wong. ML., et al. Proc Natl Acad Sci U S A. 2006; 103(41): 15124-9.

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