Technical Data

171963
Kit Type
TR-FRET
Tests
96
Detection Method
Fluorescent
EU Commodity Code
38220000
Shipping Temp
Blue Ice
Storage Temp
-20°C/-70°C
PDE1C TR-FRET BioAssay™ Kit

Phosphodiesterases (PDEs) play an important role in dynamic regulation of cAMP and cGMP signaling. PDE1C is a calmodulin-dependent PDE that is expressed principally in human myocardium.

The PDE1C TR-FRET Assay Kit is designed for identification of inhibitors of PDE1C using TR-FRET (Time Resolved Fluorescence Resonance Energy Transfer) technology. The assay is based on the generation of FAM-labeled nucleotide monophosphates by the PDE1C. 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. The PDE1C TR-FRET Assay Kit comes in a convenient 96-well format, with purified PDE1C enzyme, fluorescently labeled PDE1C substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions. Using this kit, only two simple steps on a microtiter plate are required for the PDE1C activity assay. First, the fluorescent-labeled cAMP is incubated with a sample containing PDE1C 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.
Applications
Great for screening small molecular inhibitors for drug discovery and HTS applications.
Components
PDE1C recombinant enzyme, 1x1ug FAM-Cyclic-3’, 5’-AMP: 20um, 1x20ul PDE assay buffer, 1x25ml Tb donor, 1x30ul Binding Agent, 1x200ul Binding Buffer A, 1x20ml Binding Buffer B, 1x20ml Black, low binding NUNC microtiter plate, 1 plate
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Vandeput. F., et al. J. Biol. Chem. 2007; 282(45): 32749-32757.
USBio References
No references available
United States Biological | 4 Technology Way | Salem, MA 01970
Phone 800-520-3011 | Fax 978-594-8052 | Website www.usbio.net