PDE9A2 Assay Kit

Catalog #
60381
$405 *
Size: 96 reactions
Qty
*US Pricing only. For international pricing, please contact your local distributor.
Purchase
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.

Synonyms
inhibitor screening, assay kit, PDE9A, PDE9-A2, PDE
Product Info
Storage and Usage
Citations
Assay Kit Format
Fluorescence Polarization
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.