Recombinant HEK293 cell line expressing rat PDE7A (phosphodiesterase 7A, accession number NM_031080).
N-terminal FLAG-tagged rat PDE7A has been stably expressed in a human embryonic kidney (HEK293) cell line and its expression was confirmed by Western blotting. The regulation of intracellular level of cAMP by rat PDE7A in rat PDE7A stably-expressed HEK293 cells was characterized by a cell-based reporter assay using pCRE-luc reporter vector. pCRE-luc contains a luciferase gene that is under the control of the cAMP response element (CRE). When cells transiently transfected with pCRE-luc reporter were activated by forskolin, the level of cAMP was upregulated in parental HEK293 cells inducing the expression of luciferase reporter, whereas rat PDE7A-HEK293 cells showed reduction in the level of cAMP, resulting in lowered levels of luciferase expression. Inhibition of PDE7A activity by BRL 50481, a PDE7A inhibitor, restored the cAMP level, resulting in higher luciferase activity.
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Synonyms
PDE7A, phosphodiesterase 7A, PDE7-A, PDE
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Product Data Gallery
PDE7A overexpression reduced the level of cAMP following forskolin stimulation in rat PDE7A-HEK293 cells. This effect was reversed by BRL 50481, a PDE7A inhibitor
Each vial contains ~1 X 106 cells in 1 ml of 10% DMSO.
Format
aqueous solution containing DMSO
Genbank #
NM_031080
UniProt #
O08593
Mycoplasma Testing
The cell line has been screened using the PCR-based VenorGeM Mycoplasma Detection kit (Sigma Aldrich) to confirm the absence of Mycoplasma species.
Background
Phosphodiesterases (PDEs) regulate the intracellular levels of cAMP and cGMP by
hydrolyzing cAMP and cGMP to their inactive 5' monophosphates. These cyclic
nucleotides play an important role as second messengers in diverse physiological
functions. PDE7 is a cAMP-specific enzyme and two PDE genes (PDE7A and PDE7B)
have been identified. PDE7 is widely expressed in various tissues, with PDE7A found
primarily in skeletal muscle, T lymphocytes, and pancreas, while high levels of PDE7B
are detected in brain, heart, and liver. Inhibition of PDE7 activity by its inhibitors leads to
elevated intracellular level of cAMP.
References
1. Malik, R. et al. (2008) Appl. Microbiol. Biotechnol. 77 (5): 1167-1173. 2. Fan Chung, K. (2006). Eur. J. Pharmacol. 533(1-3):110-117.
Storage/Stability
Store in liquid nitrogen immediately upon receipt.
Instructions for Use
See data sheet for detailed culturing and assay protocol.
Applications
Useful to monitor cAMP and PDE7A signaling pathway activity and to screen activators or inhibitors of PDE7A.
Shipping Temperature
-80°C
Notes
License Disclosure Visit bpsbioscience.com/license for the label license and other key information about this product.
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