NRF2/ARE Responsive Luciferase Reporter HepG2 Cell Line is derived from Human Liver cancer, and stably express firefly luciferase reporter gene under the control of NRF2/ARE response element. This cell line is an ideal cellular model for monitoring the activation of Antioxidant response Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.
NRF2 plays a crucial role in cellular anti-oxidant defense, making it a therapeutic target for neurodegenerative diseases and cancer. Under normal conditions, NRF2 localizes in the cytosol and is rapidly degraded by the proteasome. Under oxidative stress, NRF2 is stabilized and translocates to the nucleus where it binds to a DNA promoter and initiates gene expression. In the nucleus, NRF2 forms a heterodimer with a small Maf protein and binds to the Antioxidant Response Element in the upstream promoter region of many antioxidative genes, and initiates their transcription.
This NRF2 luciferase reporter HepG2 stable cell line has been stably transfected with pTA-ARE-luciferase reporter vector, which contains 4 repeats of antioxidant response binding sites, a minimal promoter upstream of the firefly luciferase coding region, along with a hygromycin expression vector. Following selection, the hygromycin resistant clones were subsequently screened for TBHQ-induced luciferase activity. The clone with the highest fold induction was selected and expanded to produce this stable cell line.
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.
