PDE1B TR-FRET Assay Kit

Catalog #: 60704
Size: 96 reactions
$350
Background:
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 assay is based on the generation of FAM-labeled nucleotide monophosphates by the phosphodiesterase. 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.
Description:
The PDE1B TR-FRET Assay Kit is designed for identification of inhibitors of PDE1B using TR-FRET (Time Resolved Fluorescence Resonance Energy Transfer) technology. 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.
Synonym(s):
inhibitor screening, FRET assay kit, PDE1B, HTRF
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.
Contraindications:
DMSO >1%, strong acids or bases, ionic detergents, high salt
Format:
Instructions for use:
See assay kit data sheet for detailed protocol.
Storage / Stability:

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.

Application(s):
Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Reference(s):
Siuciak JA, et al. (2007) Neuropharmacology 53(1):113-24.
Warning(s):
Avoid freeze/thaw cycles
Scientific Category:
PDE
Product Type:
Assay Kit
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Product Datasheets

60704.pdf