The AP1 Reporter Kit is designed for monitoring the activity of the JNK signaling pathway and the transcriptional activity of AP1 in cultured cells. The kit contains a transfection-ready AP1 luciferase reporter vector. This reporter contains the firefly luciferase gene under the control of multimerized AP1 responsive elements located upstream of a minimal promoter.
The AP1 reporter is premixed with a 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 constitutively-expressing Renilla luciferase vector as a negative control. The non-inducible luciferase vector contains the firefly luciferase gene under the control of a minimal promoter, without any additional response elements. The negative control is critical for determining pathway-specific effects and the background luciferase activity.
For assays in more difficult to transfect cell types, we suggest using our lentiviral products. The corresponding lentiviral product for this kit is BPS Bioscience #79823.
Both the Reporter and Negative Control Reporter are provided at 60 ng DNA/µl (500 µl each)
Background
The stress-activated protein kinase/c-jun N-terminal kinase (SAPK/JNK) family of proteins includes mitogen-activated protein kinases (MAPKs) that are activated by stress, inflammatory cytokines, mitogens, oncogenes, and inducers of cell differentiation and morphogenesis. Upon activation of the SAPK/JNK pathway, MAP Kinase Kinases phosphorylate and activate JNKs. The activated JNKs translocate to the nucleus where they phosphorylate and activate transcription factors such as c-Jun. The activated c-Jun forms homodimers or heterodimers with fos family proteins which bind to the activator protein-1(AP1) response element and induce target gene transcription.
References
1. Zhou H. et al. (2005) Frequency and distribution of AP-1 sites in the human genome. DNA Research. 11: 139-150. 2. Gaillard P. et.al. (2005) Design and synthesis of the first generation of novel potent, selective, and in vivo active (benzothiazol-2-yl)acetonitrile inhibitors of the c-Jun N-terminal kinase. J Med Chem. 48(14):4596-4607.
Storage/Stability
Stable at least 12 months from date of receipt, when stored as directed (-20°C)
Instructions for Use
See data sheet for detailed transfection and assay protocols.
Applications
• Monitor cAMP/PKA signaling pathway activity. • Screen for 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.
Shipping Temperature
-20°C
Notes
The vectors are suitable for transient transfection. They are NOT suitable for transformation and amplification in bacteria.
Warnings
Avoid freeze/thaw cycles
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.