PDE4D3 TR-FRET Assay Kit

Catalog #
60701
$405 *
Size: 96 reactions
Qty
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Description

Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE4D is a regulator of airway smooth-muscle contractility, and has been identified as a potential risk predictor for ischemic stroke. Additionally, PDE4D has been associated with asthma pathophysiology and bone formation. The PDE4D gene encodes at least 9 different isoforms.  

The PDE4D3 TR-FRET Assay Kit is designed for identification of inhibitors of PDE4D3 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 PDE4D3 activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE4D3 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 PDE4D3? Check out our Phosphodiesterase Screening Services.

Synonyms
inhibitor screening, FRET assay kit, PDE4D3, HTRF, PDE4-D3, PDE
Product Info
Storage and Usage
Citations
Assay Kit Format
TR-FRET
Species
Human
Supplied As
The PDE4D3 TR-FRET Assay Kit comes in a convenient 96-well format, with purified PDE4D3 enzyme, fluorescently labeled PDE substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.
Format
Catalog # Component Amount  Storage
60046 PDE4D3 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 plate 1 Room
Temp.
UniProt #
Q08499
References

Bender. A., et al. Pharmacol. Rev. 2006; 58: 488-520.

Application Reference(s):
1. Elucidating the Roles of Metabolic and Pigment-Modulating Factors in Melanoma (2019)