PDE1B Assay Kit

Catalog #
60311
$405 *
Size: 96 reactions
Qty
*US Pricing only. For international pricing, please contact your local distributor.
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Description

The PDE1B Assay Kit is designed for identification of PDE1B inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE1B to the binding agent. The key to the PDE1B Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE1B reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE1B 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.

 

Assay Principle

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Synonyms
inhibitor screening, assay kit, PDE1B, PDE1-B, PDE
Product Info
Storage and Usage
Citations
Assay Kit Format
Fluorescence Polarization
Supplied As
The PDE1B inhibitor screening assay kit comes in a convenient 96-well format, with purified PDE1B enzyme, fluorescently labeled PDE1B 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
Format
Catalog
Number

Component

Amount

Storage
60011 PDE1B 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.
UniProt #
Q01064
Background
Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE1B is a calcium-dependent cyclic nucleotide phosphodiesterase that is highly expressed in the striatum. It plays the physiological role in the central nervous system. 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.
References
Siuciak JA, et al. (2007) Neuropharmacology 53(1):113-24.