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.
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 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.
Customer Q&As
Q: Is this reporter cell line suitable for high-throughput screening (HTS)?
A: Yes. The NRF2/ARE luciferase reporter cell line produces a strong and reproducible signal and is fully compatible with 96- and 384-well HTS formats.
Q: Does this reporter respond specifically to NRF2 activation rather than general stress?
A: Yes. The reporter is designed to respond primarily to NRF2 nuclear translocation and ARE binding. Functional validation is performed using known NRF2 activators to confirm pathway-specific induction.
Q: What is the recommended treatment time for NRF2 activation assays?
A: NRF2 activation is commonly detected after 16–24 hours of compound treatment, depending on the mechanism of action of the test compound.
Q: Is the cell line compatible with co-treatment or combination studies?
A: Yes. The stable reporter system is compatible with co-treatment, time-course, and combination compound studies.
Q: Does serum concentration affect NRF2 reporter activation?
A: Serum conditions can influence basal NRF2 activity. Assay recommendations include optimized serum concentrations to minimize background and maximize induction.
Q: Can this cell line be used for environmental or chemical safety screening?
A: Yes. NRF2/ARE reporter assays are widely used for toxicity, oxidative stress, and environmental chemical screening, and this cell line is suitable for those applications.
