PDE4D (Rat) Assay Kit

Catalog #

79629

$425 *

Size: 96 Reactions

Qty

Bulk/Custom

Add to Favorites

*US Pricing only. For international pricing, please contact your local distributor.

●

Purchase

●

Description

Phosphodiesterase’s (PDEs) play an important role in the dynamic regulation of cAMP and cAMP signaling. Rat PDE4D, also known as cAMP-inhibited phosphodiesterase, has been implicated in cardiovascular function and fertility.

The Rat PDE4D Assay Kit is designed for identification of inhibitors of Rat PDE4D using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by Rat PDE4D 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 linearlypolarized 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 Rat PDE4D Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for Rat PDE4D reactions. First, the fluorescently labeled cAMP is incubated with a sample containing Rat PDE4D 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 PDE4D (Rat)? Check out our Phosphodiesterase Screening Services.

●

Synonyms

Rat PDE4D, cAMP-inhibited phosphodiesterase, cAMP-specific 3',5'-cyclic phosphodiesterase 4D, PDE4-D, PDE

●

Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

Fluorescence Polarization

Supplied As

The Rat PDE4D Assay Kit comes in a convenient 96-well format, with purified Rat PDE4D 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

Catalog #	Component	Amount	Storage
60054	Rat PDE4D 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
60390	Binding Agent	100 µl	+4°C
60391	Binding Agent Diluent (cAMP)	10 ml	+4°C
79685	Black, low binding, microtiter plate	1	Room temp.

References

1. Clapcote, S.J. Adv Neurobiol. 2017;17:103-131

2. Braun, N.N., et al., NeuroReport. 2007; Nov; 18(17):1841-1844.

Storage/Stability

Stable at least 6 months from date of receipt when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.

Instructions for Use

See assay kit data sheet for detailed 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.  

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

Phosphodiesterase

Regulatory Status

Research Use Only

Skip to the end of the images gallery

Skip to the beginning of the images gallery

PDE4D (Rat) Assay Kit

PRODUCTS

SERVICES

RESOURCES

COMPANY