The NF-κB Reporter kit is designed for monitoring the activity of the NF-κB signaling pathway in the cultured cells. The kit contains transfection-ready NF-κB luciferase reporter vector. This reporter contains a firefly luciferase gene under the control of multimerized NF-κB responsive element located upstream of a minimal promoter. The NF-κB reporter is premixed with constitutively-expressing Renilla (sea pansy) luciferase vector, which 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 a firefly luciferase gene under the control of a minimal promoter, without any additional response elements. The negative control is critical to determining pathway specific effects and 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 #79564.
Both the Reporter and Negative Control Reporter are provided at 60 ng DNA/µl (500 µl each)
Background
NF-κB (Nuclear Factor-KappaB, NFkB) is a heterodimeric protein composed of different combinations of members of the Rel family of transcription factors. The NF-kB /Rel family of transcription factors (p50, p65, c-Rel, etc.) are involved in stress, immune, and inflammatory responses. In unstimulated cells, the NF-κB dimers are sequestered in the cytoplasm by inhibitory IkB proteins. Proinflammatory cytokines, LPS, growth factors, and antigen receptors activate I B kinase (IKK), which phosphorylates the I B proteins. Phosphorylation of IkB leads to its degradation, freeing NF-κB complexes to translocate to the nucleus, bind to NF-κB DNA response elements, and induce the transcription of the target genes.
Stable at least 12 months from date of receipt, when stored as directed (-20°C)
Instructions for Use
See data sheet insert for detailed transfection and assay protocols.
Applications
• Monitor NF-κB signaling pathway activity. • Screen for activators or inhibitors of NF-κB signaling or NF-κB pathway components. • Study effects of RNAi or gene overexpression on the activity of the NF-κB 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.
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