PDE4D7 TR-FRET Assay Kit

Catalog #

60708

$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. 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.

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Synonyms

inhibitor screening, FRET assay kit, PDE4D7, HTRF, PDE4-D7, PDE

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

TR-FRET

Species

Human

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.

Storage/Stability

At least 6 months from date of receipt when stored as directed.

Applications

Great for screening small molecular inhibitors for drug discovery and HTS applications.

Shipping Temperature

-80°C

Warnings

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

Regulatory Status

Research Use Only