PDE1B TR-FRET Assay Kit

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.

DETAILS

Shiptemp: -80°C (dry ice)
Synonyms: inhibitor screening, FRET assay kit, PDE1B
Warnings: Avoid freeze/thaw cycles.
Category: Phosphodiesterase/Assay Kit
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.
References: Siuciak JA, et al. (2007) Neuropharmacology 53(1):113-24.
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.
Methodology: 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.
Unspsc Code: 41121811
Unspsc Name: Laboratory microchemistry kits
Applications: Great for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Product Type: Assay Kit
Biosafety Level: Not applicable (BSL-1)
Contraindications: DMSO >1%, strong acids or bases, ionic detergents, high salt
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.
Scientific Category: Phosphodiesterase
Instructions for Use: See assay kit datasheet for detailed protocol.