Product Name Catalog # Price (NP)**   Qty
NRF2/ARE Luciferase Reporter MCF7 Stable Cell Line SL-0010-NP $1,200
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  • ** Non Profit (NP) price is for academic, non profit organizations and institutes
Description:

NRF2/ARE Responsive Luciferase Reporter MCF7 Stable Cell Line is derived from human breast cancer, and stably express firefly luciferase reporter gene under the control of the NRF2/ARE response element.  This cell line is an ideal cellular model for monitoring the activation of Antioxidant Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.


Principle

NRF2 is a transcription factor, which plays an important role in responding to oxidative stress. Under normal cellular conditions, NRF2 forms a protein complex with Keap1 in the cytoplasm, which results in the proteasomal degradation of NRF2 causing it to be inactive.  Oxidative stress leads to the activation of a number of kinases including MAPK, ERK, p38, PKC, and PI3K.  They phosphorylate both Keap1 and NRF2, which disrupt the Keap1-NRF2 complex, and stimulate the translocation of NRF2 to the nucleus, where it forms a complex with Maf proteins.  The NRF2/Maf heterodimers bind directly to antioxidant response elements (AREs) located within promoters of NRF2 target genes and coordinate the expression of antioxidant gene products. 

Signosis has developed a NRF2-reporter MCF7 stable cell line that has been stably transfected with pTA-NRF2-luciferase reporter vector, which contains 8 repeats of NRF2 binding sites, a minimal promoter upstream of the firefly luciferase coding region, along with a G418 expression vector.  This cell line can be used to investigate oxidative stress-mediated activation of upstream kinases and to screen anticancer drugs that can induce ARE-driven gene expression.

 

Principle behind TF luciferase reporter.  TF luciferase reporter stable cell line utilizes artificial promoter constructs to drive luciferase expression.  The promoter region can consists of multiple repeats of a cis-element TF binding site, a DNA fragment from the promoter region of a known TF downstream gene, or a DNA fragment containing putative/known TF binding sites.  There are several ways that a TF can be activated, such as through extracellular stimuli or through intracellular signaling pathways.  Once activated, the TF translocates to the nucleus and often interacts with relevant co-factors to drive gene expression.  Once luciferase is expressed, it can generate light in an enzymatic assay and the amount of light measured is positively correlated with the level of TF activation.

 

Data

Analysis of the NRF2 Pathway Reporter MCF7 Stable Cell Line in response to stimuli. The cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were treated with or without 50nM TBHQ respectively in DMEM and 0.1% FBS for 16 hours.

Literature

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