PDE4D2 Assay Kit

Catalog #: 60345
Size: 96 reactions
$350
Background:
Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE4D is a regulator of airway smooth-muscle contractility, and has been identified as a potential risk predictor for ischemic stroke. Additionally, PDE4D has been associated with asthma pathophysiology and bone formation. The PDE4D gene encodes at least 9 different isoforms. 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 PDE4D2 Assay Kit is designed for identification of inhibitors of PDE4D2 using fluorescence polarization. The assay is based on the binding of a fluorescent nucleotide monophosphate generated by PDE4D2 to the binding agent. The key to the PDE4D2 Assay Kit is the specific binding agent. Using this kit, only two simple steps on a microtiter plate are required for PDE4D2 reactions. First, the fluorescently labeled cAMP is incubated with a sample containing PDE4D2 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 equipped for the measurement of fluorescence polarization.
Synonym(s):
inhibitor screening, assay kit, PDE4D
Supplied As:
The PDE4D2 Assay Kit comes in a convenient 96-well format, with purified PDE4D2enzyme, fluorescently labeled PDE4D substrate (cAMP), binding agent, and PDE assaybuffer for 100 enzyme reactions.
Contraindications:
DMSO >1%, strong acids or bases, ionic detergents, high salt
Format:

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.
Notes:
MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:
Fluorescent microplate reader capable to measure fluorescence polarization

License Disclosure: Purchase of this cell line grants you with a 10-year license to use this cell line in your immediate laboratory, for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make the cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit the use of this cell line in humans or for therapeutic or drug use. Inappropriate use or distribution of this cell line will result in revocation of the license and result in an immediate cease of sales and distribution of BPS products to your laboratory. Publications using this cell line should reference BPS Bioscience, Inc., San Diego.

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.

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Product Datasheets

60345.pdf