PDE10A2 TR-FRET Assay Kit

Catalog #
60710
$405 *
Size: 96 reactions
Qty
*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. PDE10A is a dual substrate PDE highly expressed in striatal medium spiny neurons. PDE10A inhibitors can improve the cognitive symptoms of schizophrenia, and exhibit potential therapeutic value for Huntington’s disease. PDE10A2 is located in cytosol, whereas PDE10A2 is a membrane-associated protein.

The PDE10A2 TR-FRET Assay Kit is designed for identification of inhibitors of PDE10A2 using TR-FRET (Time Resolved Fluorescence Resonance Energy Transfer) technology. The assay is based on the generation of FAM-labeled nucleotide monophosphates by the phosphodiesterase. These phosphate groups bind to terbium-labeled nanoparticles, resulting in energy transfer from the terbium to the FAM, which emits a fluorescent signal at 520 nm. The change in fluorescent intensity can be easily measured using a fluorescence plate reader.

Using this kit, only two simple steps on a microtiter plate are required for the PDE10A2 activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE10A2 for 1 hour. Second, a binding agent and a terbium donor are added to the reaction mix and incubated for 1 hour. Then, fluorescence intensity can be measured using a fluorescence reader.

Need us to run inhibitor screens or profile your compounds against PDE10A2? Check out our Phosphodiesterase Screening Services.

Synonyms
inhibitor screening, FRET assay kit, PDE10A2, HTRF, PDE10-A2, PDE
Product Info
Storage and Usage
Citations
Assay Kit Format
TR-FRET
Species
Human
Supplied As
The PDE10A2 TR-FRET Assay Kit comes in a convenient 96-well format, with purified PDE10A2 enzyme, fluorescently labeled PDE substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
Materials Required But Not Supplied

Fluorescent microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET)

Format
Catalog # Component Amount Storage 
60100 PDE10A2 recombinant enzyme 1 µg -80°C Avoid
freeze/
thaw
cycles!
60200 FAM-Cyclic-3′, 5′-AMP: 20 µM 50 µl -80°C
60393 PDE assay buffer 25 ml -20°C
60394 Tb donor 50 µl -80°C
60390 Binding Agent 200 µl +4°C
78422 Binding Buffer A 20 ml +4°C
78423 Binding Buffer B 20 ml +4°C
79685 Black, low binding NUNC microtiter pla 1 Room Temp.
UniProt #
Q9Y233
References

1. Akinori Nishi and Gretchen L. Snyder. J Pharmacol Sci., 2010, 114: 6 – 16.
2. Kenji Omori and Jun Kotera. Circulation Research. 2007; 100: 309-327.