PDE9A2 Assay Kit

Catalog #

60381

$405 *

Size: 96 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE9A2 is primarily expressed in spleen, small intestine, and brain. PDE9 inhibitors have been studied as therapeutics for treatment of cardiovascular diseases, diabetes, and neurodegenerative disorders. The PDE9A2 Assay Kit is designed for identification of PDE9A2 inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE9A2 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 linearlypolarized 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 Fluorescence Polarization signal is measured using a fluorescent microplate reader capable of measuring fluorescence polarization.

Need us to run inhibitor screens or profile your compounds against PDE9A2? Check out our Phosphodiesterase Screening Services.

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Synonyms

inhibitor screening, assay kit, PDE9A, PDE9-A2, PDE

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

Fluorescence Polarization

Species

Human

Supplied As

The PDE9A inhibitor screening assay kit comes in a convenient 96-well format, includingpurified PDE9A enzyme, fluorescently labeled PDE9 substrate (cGMP), binding agent,and PDE assay buffer for 100 enzyme reactions.

Materials Required But Not Supplied

Fluorescent microplate reader capable to measure fluorescence polarization.
Adjustable micropipettor and sterile tips.
Inhibitor Buffer (inhibitor solution without inhibitor)

Format

Catalog Number	Component	Amount	Storage
60090	PDE9A2 recombinant enzyme*	>1 µg	-80°C	Avoid freeze/ thaw cycles!
60201	FAM-Cyclic-3′, 5′-GMP( 20 µM)	50 µl	-80°C
60393	PDE assay buffer	25 ml	-20°C
60390	Binding Agent	100 µl	+4°C
60392	Binding Agent Diluent (cGMP)	10 ml	+4°C
79685	Black, low binding, microtiter plate	1	Room temp.

*We have provided additional material in this vial for ease of retrieval.

UniProt #

O76083

Background

Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE9A is primarily expressed in spleen, small intestine, and brain. PDE9 inhibitors have been studied as therapeutics for treatment of cardiovascular diseases, diabetes, and neurodegenerative disorders. 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.

References

Wang, H. et al. (2010) J. Med Chem. 53 (4): 1726-31.

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

Contraindications

DMSO >1%, strong acids or bases, ionic detergents, high salt

Warnings

This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to 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.

Scientific Category

PDE, Neuroscience

Regulatory Status

Research Use Only