Transfection Collection(TM) : SRE Transient Pack (MAPK/ERK Signaling Pathway)

The SRE Reporter Kit Transient Pack is designed to provide the tools necessary for transiently transfecting and monitoring the activity of the ERK signaling pathway and the transcriptional activity of SRE in cultured HEK293 cells. The kit contains transfection-ready vectors containing firefly luciferase as a MAPK/ERK 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 SRE Transient Pack is the SRE luciferase reporter vector, which is an ERK pathway-responsive reporter. This reporter contains the firefly luciferase gene under the control of multimerized SRE responsive elements located upstream of a minimal promoter. The SRE reporter is premixed with a constitutively-expressing Renilla luciferase vector that 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 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. 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 luciferase 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. * Other mammalian cell lines may also be used, but an alternate cell culture medium may be required for optimal cell growth.