PDE4D7 TR-FRET Assay Kit

Catalog #
60708
$405 *
Size: 96 reactions
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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 PDE4D7 TR-FRET Assay Kit is designed for identification of inhibitors of PDE4D7 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 PDE4D7 activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE4D7 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.

Synonyms
inhibitor screening, FRET assay kit, PDE4D7, HTRF, PDE4-D7, PDE
Product Info
Storage and Usage
Citations
Assay Kit Format
TR-FRET
Supplied As
The PDE4D7 TR-FRET Assay Kit comes in a convenient 96-well format, with purified PDE4D7 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
60047 PDE4D7 recombinant enzyme 1 µg -80°C Avoid freeze/ thaw cycles
60200 FAM-Cyclic-3′, 5′-AMP: 20 µM 50 µl -20°C
60393 PDE assay buffer 25 ml -80°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 microtiter plate 1 Room Temp
UniProt #
Q08499
References
Bender. A., et al. Pharmacol. Rev. 2006; 58: 488-520.