PDE4D Cell-Based Activity Assay Kit
Catalog #: 60505

Size: 500 reactions

Description
Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP
and cGMP signaling. PDE4D has 3',5'-cyclic-AMP phosphodiesterase activity and
degrades cAMP. Inhibition of PDE4D activity by its inhibitors leads to an elevated
intracellular level of cAMP. The PDE4D gene encodes at least 9 different isoforms, and
has been linked to stroke, asthma, arrhythmia, and cardiac myopathy, making it an
important therapeutic target.

The PDE4D cell-based activity assay is designed for screening inhibitors of PDE4D7 in
cultured 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 response
element (CRE). Elevation of intracellular cAMP activates CRE binding protein (CREB)
to bind CRE and induce the expression of luciferase. Forskolin is commonly used to
raise the intracellular level of cAMP in cell physiology studies. When cells transiently
transfected with CRE reporter are activated by forskolin, the intracellular level of cAMP is
upregulated, which induces the expression of CRE luciferase reporter. However, when
cells are co-transfected with PDE4D7 expression vector and CRE reporter, the level of
forskolin-induced cAMP is reduced, resulting in lower expression level of luciferase.
When cells are treated with PDE4D inhibitor to inhibit PDE4D7 activity, cAMP level is
restored, 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.

Components
 

These vectors are ready for transient transfection. They are NOT MEANT for
transformation and amplication in bacteria.

References
Montminy MR et al. (1987) Binding of a nuclear protein to the cyclic-AMP response
element 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 and
development of an assay for screening of PDE7 selective inhibitors. Appl.
Microbiol. Biotechnol. 77 (5): 1167-1173