A 96-well assay for measurement of Nrf2.

• A sensitive, non-radioactive method of detecting Nrf2 from whole cell lysates
• 96-well plate format replaces EMSAs
• Capture the transcription factor using a specific dsDNA sequence bound to the plate
• Detect the dsDNA-bound transcription factor with specific antibodies in an ELISA format

Nuclear factor E2-related factor 2 (Nrf2) is a transcription factor that plays a key role in maintaining redox homeostasis via its interaction with a cysteine-rich protein Kelch-like ECH-associated protein 1 (Keap1). In resting cells, Nrf2 and Keap1 form a tight complex, which is targeted for degradation by proteasomes.

Under oxidative stress, Nrf2 is released from the Nrf2/Keap1 complex and translocate to the nucleus where it is able to induce the expression of a battery of genes encoding diverse cytoprotective proteins, including antioxidative enzymes, anti-inflammatory mediators, and proteasomes. Nrf2 Transcription Factor Assay is a non-radioactive, colorimetric method for detecting specific transcription factor DNA binding activity in nuclear extracts.

A specific double stranded DNA (dsDNA) sequence containing the Nrf2 response element is immobilized onto the wells of a 96-well plate. Nrf2 contained in nuclear extract samples will bind specifically to the Nrf2 response element and can be detected by addition of a specific Nrf2 antibody and a secondary antibody conjugated to HRP.