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The pNL3.2.NF-kappaB-RE[NlucP/NF-kappaB-RE/Hygro] Vector contains five copies of an NF-kappaB response element (NF-kappaB-RE) that drives transcription of a destabilized form of NanoLuc luciferase, an engineered small (23.3kDa) luciferase fusion protein. The NlucP reporter consists of NanoLuc luciferase with a C-terminal fusion to PEST, a protein destabilization domain, which responds more quickly and with greater magnitude to changes in transcriptional activity than unmodified NanoLuc luciferase. NanoLuc (Nluc) luciferase is a small enzyme (19.1kDa) engineered for optimal performance as a luminescent reporter. The enzyme is about 100-fold brighter than either firefly (Photinus pyralis) or Renilla reniformis luciferase using a novel substrate, furimazine, to produce high intensity, glow-type luminescence. The luminescent reaction is ATP-independent and designed to suppress background luminescence for maximal assay sensitivity.
