PDE3A Assay Kit

Catalog #

79736

$425 *

Size: 96 reactions

Qty

Bulk/Custom

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Description

Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE3A, also known as cGMP-inhibited phosphodiesterase, has been implicated in cardiovascular function and fertility. The PDE3A Assay Kit is designed for identification of PDE3A inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE3A 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 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.

The key to the PDE3A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE3A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE3A for 1 hour. Second, binding agent is added to the reaction mix to produce a change in fluorescent polarization that can then be measured using a fluorescence reader equipped for the measurement of fluorescence polarization.

Need us to run inhibitor screens or profile your compounds against PDE3A? Check out our Phosphodiesterase Screening Services.

This product has been cited 7 times.

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Synonyms

Phosphodiesterase 3A, PDE3A, PDE3-A, PDE

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Product Data Gallery

Product Info

Storage and Usage

Citations7

Assay Kit Format

Fluorescence Polarization

Supplied As

The PDE3A inhibitor screening assay kit comes in a convenient 96-well format, with purified human PDE3A enzyme, fluorescently labeled PDE3 substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.

Materials Required But Not Supplied

Fluorescent microplate reader capable of measuring fluorescence polarization.
Adjustable micropipettor and sterile tips.
1,4-Dithiothreitol (DTT) 1 M in anhydrous DMSO.

Format

Catalog Number	Component	Amount	Storage
60032	PDE3A recombinant enzyme	>1 µg	-80°C	*Avoid freeze/* *thaw cylces!*
60200	FAM-Cyclic-3´, 5´-AMP (20 μM)	50 µl	-80°C
60393	PDE assay buffer	25 ml	-20°C
60390	Binding Agent	100 µl	+4°C
60391	Binding Agent Diluent	10 ml	+4°C
79685	Black, low binding, microtiter plate	1	Room Temp.

UniProt #

Q14432

References

Maurice DH. Front. Biosci. 2005; 10:1221-8.

Storage/Stability

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

Instructions for Use

See data sheet for detailed assay protocol.

Shipping Temperature

-80°C

Warnings

This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to read the FP signal. For more information FP technology, visit our Tech Note: FP, assay principles and applications.  

Avoid multiple 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