CRE/CREB Reporter Assay Kit
cAMP/PKA Cell Signaling Pathway
Catalog #: 60611

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 CREB and is
responsible for its activation. CRE is at the focus 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
and 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.

Description
The CRE/CREB Reporter kit is designed for monitoring the activity of the cAMP/PKA
signaling pathway in cultured cells. The kit contains transfection-ready CRE luciferase
reporter. This reporter contains the firefly luciferase gene under the control of
multimerized cAMP response element (CRE) located upstream of a minimal promoter.
Elevation of the intracellular cAMP level activates cAMP response element binding
protein (CREB) to bind CRE and induces the expression of luciferase.

The CRE reporter is premixed with constitutively-expressing Renilla (sea pansy)
luciferase vector that serves as an internal control for transfection efficiency. The kit also
includes a non-inducible firefly luciferase vector premixed with the constitutivelyexpressing
Renilla luciferase vector as a negative control. The non-inducible luciferase
vector contains the firefly luciferase gene under the control of a minimal promoter, but
without any additional response elements. The negative control is critical to determining
pathway-specific effects and background luciferase activity.

Applications• Monitor cAMP/PKA signaling pathway activity.
• Screen activators or inhibitors of PKA or cAMP/PKA pathway components.
• Study effects of RNAi or gene overexpression on the activity of the cAMP/PKA
  pathway.

Components

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

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.