Cellular oxidative and electrophilic stress caused by drugs and other xenobiotics, inflammation, and ionizing radiation are associated with an accumulation of reactive oxygen species and electrophilic insults, which contribute to the pathogenesis of various diseases such as cancer, neurodegenerative disease, and atherosclerosis. In order to protect cells from reactive oxygen species and electrophilic insults, the endogenous cellular antioxidant defense system initiates a response to cellular oxidative and electrophilic stress. NRF2 (nuclear factor (erythroid-derived 2)-like 2; NFE2L2) is a key transcriptional factor regulating hundreds of antioxidant and Phase II detoxification genes. Under normal conditions, NRF2 is sequestered in the cytoplasm through binding with Keap1, an actin-binding protein and finally is degraded through the Keap1-dependent ubiquitination. In response to a stimulus, degradation of Keap1 is markedly increased. This leads to the disruption of the Keap1-NRF2 complex and nuclear translocation of NRF2. NRF2 then dimerizes with small Maf proteins and binds to the ARE (antioxidative response element) in promoters of downstream genes to initiate expression.

• Specific transcription factor-DNA binding assay
• Perfect alternative to EMSA
• Easy to perform in an ELISA format
• Non-radioactive assay
• High throughput (96-well plate format)
• Assay can be completed within 5 hours

Upon receipt, the positive control should be removed and stored at -20^° or -80^°C. The remainder of the kit can be stored for up to 6 months at 2-8^°C from the date of shipment. Opened Microplate Wells or reagents may be stored for up to 1 month at 2^° to 8^°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge.
Note: The kit can be used within one year if the whole kit is stored at -20^°C upon receipt. Avoid repeated freeze-thaw cycles.