PDE5A1 Assay Kit

Catalog #: 60350
Size: 96 reactions
$350
Background:
Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE5A1, also known as cGMP-inhibited phosphodiesterase, has been implicated in cardiovascular function and fertility. 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.
Description:
The PDE5A1 Assay Kit is designed for identification of PDE5A1 inhibitors using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE5A1 to the binding agent. The key to the PDE5A1 Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE5A1 reactions. First, the fluorescently labeled cGMP is incubated with PDE5A1 for 1 hour. Second, the binding agent is added to the reaction mix to produce a change in fluorescent polarization that can then be measured using a fluorescence reader equipped for the measurement of fluorescence polarization.
Materials Required But Not Provided:
Fluorescent microplate reader capable to measure fluorescence polarization
Synonym(s):
inhibitor screening, assay kit, PDE5
Supplied As:
The PDE5A1 inhibitor screening assay kit comes in a convenient 96-well format, withpurified PDE5A1 enzyme, fluorescently labeled PDE5 substrate (cGMP), binding agent,and PDE assay buffer for 100 enzyme reactions.
Contraindications:
DMSO >1%, strong acids or bases, ionic detergents, high salt
Format:

Assay Kit

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):
Warning(s):
Avoid freeze/thaw cycles
Scientific Category:
PDE
Product Type:
Assay Kit
Data shown is lot-specific. Contact us for specific information on other lots.

Quality Assurance

Product Datasheets

60350.pdf