PDE3A (Dog) Assay Kit

Catalog #

79735

$495 *

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 the dynamic regulation of cAMP and cAMP signaling. Dog PDE3A, also known as cGMP-inhibited phosphodiesterase, has been implicated in cardiovascular function and fertility. The Dog PDE3A Assay Kit is designed for identification of inhibitors of canine PDE3A using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by Dog 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 Dog PDE3A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for Dog PDE3A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing Dog PDE3A for 1 hour. Second, a 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 (Dog)? Check out our Phosphodiesterase Screening Services.

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Synonyms

Phosphodiesterase 3A, PDE3A, PDE3-A, PDE

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

Fluorescence Polarization

Supplied As

The Dog PDE3A Assay Kit comes in a convenient 96-well format, with purified Dog PDE3A1 enzyme, fluorescently labeled substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions.

Materials Required But Not Supplied

Fluorescent microplate reader capable to measure fluorescence polarization.

Adjustable micropipettor and sterile tips.

1,4-Dithiothreitol (DTT) 1 M in anhydrous DMSO.

Format

Assay Kit Components

References

1. Hung, S.H., et al. J. Biol. Chem. 281 (39), 29236-29244 (2006)
2. Hambleton, R., et al. J. Biol. Chem. 280 (47), 39168-39174 (2005)

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

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PDE3A (Dog) Assay Kit

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