NF-kappaB Dual Reporter Vector

The NF-kB signaling pathway (Nuclear Factor kappa-light-chain-enhancer of activated B cells, a family of transcription factors regulating immune and inflammatory responses) coordinates immune responses, inflammation, cell survival, and proliferation, with dysregulation linked to chronic inflammation, autoimmunity, and cancer. NF-kB transcription factors are activated by diverse stimuli (TNF-alpha, IL-1beta, LPS) through upstream kinases (IKK complex), leading to IkB degradation, NF-kB nuclear translocation, and target gene expression. The NF-kB Dual Reporter Vector (NKRV) is a DNA-based dual-luciferase system engineered to monitor this NF-kB-dependent transcriptional activity in mammalian cells. It comprises an NF-kB Reporter Mix, in which Firefly luciferase is driven by multimerized NF-kB response elements and co-formulated with a constitutive Renilla luciferase plasmid for internal normalization, and a Negative Control Reporter Mix lacking NF-kB binding sites to define non-inducible background and pathway specificity. Firefly luciferase provides a strong ATP-dependent signal proportional to NF-kB-driven transcription, while Renilla luciferase offers an ATP-independent internal control, enabling accurate quantification of NF-kB pathway activity suitable for pathway studies, genetic perturbation experiments, and screening of small-molecule or biologic modulators.