PDE1B TR-FRET Assay Kit

Catalog #
60704
$405 *
Size: 96 reactions
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Description

Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE1B is a calcium/calmodulin-dependent cyclic nucleotide phosphodiesterase that is highly expressed in the striatum. It plays a physiological role in the central nervous system, and PDE1B activity has been linked to impaired cognition and spatial learning.

The PDE1B TR-FRET Assay Kit is designed for identification of inhibitors of PDE1B using TR-FRET (Time Resolved Fluorescence Resonance Energy Transfer) technology. The assay is based on the generation of FAM-labeled nucleotide monophosphates by the PDE1B. 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 PDE1B activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE1B for 1 hour. Second, a binding agent and a terbium donor are added to the reaction mix and incubated for 30 minutes. Then, fluorescence intensity can be measured using a fluorescence reader.

Need us to run inhibitor screens or profile your compounds against PDE1B? Check out our Phosphodiesterase Screening Services.

Synonyms
inhibitor screening, FRET assay kit, PDE1B, HTRF, PDE1-B, PDE
Product Info
Storage and Usage
Citations
Assay Kit Format
TR-FRET
Supplied As
The PDE1B TR-FRET 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.
Format
Catalog # Component Amount Storage
60011 PDE1B recombinant enzyme 1 µg -80°C Avoid freeze/ thaw cycles!
60200 FAM-Cyclic-3′, 5′-AMP: 20 µM 20 µl -80°C
60393 PDE assay buffer 25 ml -20°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 #
Q01064
References
Siuciak JA, et al. (2007) Neuropharmacology 53(1):113-24.