Transfection Collection™ : NF-κB Transient Pack ( NF-κB Signaling Pathway)

The NF-κB Transient Pack is designed to provide the tools necessary for transiently transfecting and monitoring the activity of the NF-κB signaling pathway in cultured HEK293 cells. The kit contains transfection-ready vectors containing firefly luciferase as a NF-κB pathway-responsive reporter and constitutively expressing Renilla luciferase as a transfection control. It also includes the Dual Luciferase detection reagents to detect both luciferase activities and specialized medium for growing and assaying HEK293 cells. The key to the NF-κB Transient Pack is the 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 luciferase vector, which serves as an internal control for transfection efficiency. The pack 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. Additionally, the pack includes cell culture medium (BPS Medium 1) that has been optimized for use with HEK293 cells*. BPS Medium 1 includes MEM medium, 10% fetal bovine serum, 1% non-essential amino acids, sodium pyruvate, and 1% Pen/Strep. Finally, the pack provides the Dual Luciferase (Firefly-Renilla) Assay System. These reagents provide highly sensitive, stable detection of firefly luciferase activity and Renilla luciferase activity. The dual luciferase reagents can be used directly in cells in growth medium, and can be detected with any luminometer; automated injectors are not required. *Note: the kit may be used with other cell lines than HEK293, but an alternate cell culture medium may be required for optimal cell growth.

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