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Applications: Monitor cAMP/PKA signaling pathway activity.
Screen for activators or inhibitors of the cAMP/PKA signaling pathway.
Background: The main role of the cAMP response element, or CRE, is mediating the effects of Protein Kinase A (PKA) by way of transcription. It is the main binding site of cAMP response element binding protein (CREB) and is responsible for its activation. CRE is the target of many extracellular and intracellular signaling pathways, including cAMP, calcium, GPCR (G-protein coupled receptors) and neurotrophins. The cAMP/PKA signaling pathway is critical to numerous life processes in living organisms. In the cAMP/PKA signaling pathway, CREB is activated via phosphorylation of PKA and binds to CRE with a general motif of 5'-TGACGTCA-3'. Since CRE is a modulator of the cAMP/PKA signaling pathway, it allows the effects of various inhibitors to be studied.
Biological Activity: This cell line is validated for response to stimulation by Forskolin and to the treatment with an inhibitor of the cAMP/PKA signaling pathway.
Description: The cAMP/PKA Signaling Pathway CRE/CREB Reporter (Luc) - HEK293 Cell Line is designed for monitoring the activity of the cAMP/ PKA signaling pathway. The cAMP/PKA Signaling Pathway CRE/CREB Reporter (Luc) - HEK293 Cell Line contains a firefly luciferase gene under the control of multimerized cAMP response element (CRE) stably integrated into HEK293 cells. Elevation of the intracellular cAMP level activates cAMP response element binding protein (CREB) to bind CRE and induces the expression of luciferase.
Format: aqueous solution containing DMSO
Host Cell Line: HEK293
Mycoplasma Testing: The cell line has been screened using the PCR-based Venor™GeM Mycoplasma Detection kit (Sigma Aldrich) to confirm the absence of Mycoplasma species.
Storage Stability: Store in liquid nitrogen immediately upon receipt.
Supplied As: Each vial contains ~1.5 X 10^6 cells in 1 ml of 10% DMSO.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: 1. Montminy, M.R. et al. (1987) Binding of a nuclear protein to the cyclic-AMP response element of the somatostatin gene. Nature 328(6126): 175-178.
2. Fan Chung, K. (2006) Phosphodiesterase inhibitors in airways disease. Eur. J. Pharmacol. 533(1-3): 110-117.
3. 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.
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