PDE4D Cell-Based Activity Assay Kit / BPS Bioscience Product-Datasheet BPS-60505
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PDE4D Cell-Based Activity Assay Kit
PDE4D Cell-Based Activity Assay KitCatalog #: BPS-60505
Size: 500 reactions
DescriptionPhosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMPand cGMP signaling. PDE4D has 3',5'-cyclic-AMP phosphodiesterase activity anddegrades cAMP. Inhibition of PDE4D activity by its inhibitors leads to an elevatedintracellular level of cAMP. The PDE4D gene encodes at least 9 different isoforms, andhas been linked to stroke, asthma, arrhythmia, and cardiac myopathy, making it animportant therapeutic target.
The PDE4D cell-based activity assay is designed for screening inhibitors of PDE4D7 incultured cells. The assay is based on transfecting cells with the CRE luciferase reporter.CRE reporter contains the firefly luciferase gene under the control of cAMP responseelement (CRE). Elevation of intracellular cAMP activates CRE binding protein (CREB)to bind CRE and induce the expression of luciferase. Forskolin is commonly used toraise the intracellular level of cAMP in cell physiology studies. When cells transientlytransfected with CRE reporter are activated by forskolin, the intracellular level of cAMP isupregulated, which induces the expression of CRE luciferase reporter. However, whencells are co-transfected with PDE4D7 expression vector and CRE reporter, the level offorskolin-induced cAMP is reduced, resulting in lower expression level of luciferase.When cells are treated with PDE4D inhibitor to inhibit PDE4D7 activity, cAMP level isrestored, resulting in higher luciferase activity.
The kit includes CRE luciferase reporter (premixed with constitutively-expressing Renilla(sea pansy) luciferase vector that serves as an internal control for transfection efficiency),PDE4D7 expression vector, and forskolin.
Applications• Screen PDE4D inhibitors for drug discovery.• Monitor cAMP/PDE4D signaling pathway activity.
These vectors are ready for transient transfection. They are NOT MEANT fortransformation and amplication in bacteria.
ReferencesMontminy MR et al. (1987) Binding of a nuclear protein to the cyclic-AMP responseelement of the somatostatin gene. Nature 328(6126):175-178
Fan Chung, K. (2006) Phosphodiesterase inhibitors in airways disease. Eur. J.Pharmacol. 533(1-3):110-117
Malik, R. et al. (2008) Cloning, stable expression of human phosphodiesterase 7A anddevelopment of an assay for screening of PDE7 selective inhibitors. Appl.Microbiol. Biotechnol. 77 (5): 1167-1173
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