SRE Reporter Kit (MAPK/ERK Signaling Pathway)
The SRE Reporter Kit is designed for monitoring the activity of the ERK signaling pathway and the transcriptional activity of SRF in cultured cells. The kit contains a transfection-ready 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 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 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.
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 #78627.
This product has been cited 2 times.
Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.
Negative Control Reporter: Non-inducible luciferase vector + constitutively-expressing Renilla luciferase vector
- Mammalian cell line and appropriate cell culture medium
- 96-well tissue culture plate or 96-well tissue culture-treated white clear-bottom assay plate
- Transfection reagent for mammalian cell line [We use Lipofectamine™ 2000 (Invitrogen # 11668027). However, other transfection reagents work equally well.]
- Opti-MEM I Reduced Serum Medium (Invitrogen #31985-062)
- TWO-Step Luciferase Assay System:TWO-Step Luciferase (Firefly & Renilla) Assay System (BPS Bioscience #60683): This system assays cells directly in growth medium. It can be used with any luminometer. Automated injectors are not required.
- Luminometer
TWO-Step Luciferase (Firefly & Renilla) Assay System (BPS Bioscience #60683): This system assays cells directly in growth medium. It can be used with any luminometer. Automated injectors are not required.
Both the Reporter and Negative Control Reporter are provided at 55 ng DNA/µl (500 µl each)
The MAPK/ERK signaling pathway is a major participant in the regulation of cell growth and differentiation. It can be activated by various extracellular stimuli including mitogens, growth factors, and cytokines. Upon stimulation, MEK1/2 phosphorylate and activate ERK1/2. The activated ERK translocates to the nucleus where it phosphorylates and activates transcription factors. The TCFs (Ternary Complex Factors), including Elk1, are among the best-characterized transcription factor substrates of ERK. When phosphorylated by ERK, Elk1 forms a complex with Serum Response Factor (SRF) and binds to Serum Response Element (SRE), resulting in the expression of numerous mitogen-inducible genes.
1. Wong, K.K. (2009) Recent developments in anti-cancer agents targeting the Ras/Raf/ MEK/ERK pathway. Recent Pat Anticancer Drug Discov. 4(1):28-35.
2. Treisman, R. (1992) The serum response element. Trends Biochem Sci. 17(10): 423-426.
Application Reference(s):
1. Efficient, continuous mutagenesis in human cells using a pseudo-random DNA editor (2019)
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