PDE1C Assay Kit, BioAssay(TM)

Product Overview

• Article Name: PDE1C Assay Kit, BioAssay(TM)
• Biozol Catalog Number: USB-137330
• Supplier Catalog Number: 137330
• Alternative Catalog Number: USB-137330-96
• Manufacturer: US Biological
• Category: Kits/Assays

Product Description

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. Intended Use: The PDE1C Assay Kit is designed for identification of PDE1C inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE1C to the binding agent. 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. Test Principle: The PDE1C inhibitor screening assay kit comes in a convenient 96-well format, including purified PDE1C enzyme, fluorescently labeled PDE1C substrate (cAMP), binding agent, and PDE assay buffer for 100 enzyme reactions. The key to the PDE1C Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE1C reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE1C 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. Kit Components: NUNC microtiter plate, 1x96wells Recombinant enzyme, 1x1ug FAM-Cyclic-3,5-AMP 20uM, 1x20ul PDE assay buffer, 1x25ml Binding agent, 1x200ul Binding agent diluent, 1x20ml Materials or Instruments Required But Not Supplied: Fluorescent microplate reader capable of measuring fluorescence polarization Applications: Suitable for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications. Storage and Stability: Store other components at 4C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.