PDE1A Assay Kit

DESCRIPTION

The PDE1A Assay Kit is designed for identification of PDE1A inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE1A1 to the binding agent. The key to the PDE1A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE1A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE1A1 for 1 hour. Second, a binding agent is added to the reaction mix to produce a change in fluorescent polarization. The FP signal is measured using a fluorescent microplate reader capable of measuring fluorescence polarization.

DETAILS

Notes: MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED: Fluorescent microplate reader capable to measure fluorescence polarization
Shiptemp: -80°C (dry ice)
Synonyms: inhibitor screening, assay kit, PDE1A
Warnings: Avoid freeze/thaw cycles.
Category: Phosphodiesterase/Assay Kit
Background: Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE1A1 is a calmodulin-dependent PDE and expressed in lung and heart. PDE1A1 is believed to regulate vascular smooth muscle contraction and may play a role in sperm function. 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.
References: Bender. A., et al. Pharmacol. Rev. 2006; 58: 488-520.
Description: The PDE1A Assay Kit is designed for identification of PDE1A inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE1A1 to the binding agent. The key to the PDE1A Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE1A reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE1A1 for 1 hour. Second, a binding agent is added to the reaction mix to produce a change in fluorescent polarization. The FP signal is measured using a fluorescent microplate reader capable of measuring fluorescence polarization.
Methodology: Fluorescence Polarization
Supplied As: The PDE1A inhibitor screening assay kit comes in a convenient 96-well format, including purified PDE1A1 enzyme, fluorescently labeled PDE1A 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.