4.8. Nrf2 Activity Assay

Nrf2 activity was analyzed using the Nrf2 Transcription Factor Assay Kit (#ab207223, Abcam, Cambridge, UK) according to the manufacturer’s protocol. Briefly, nuclear extracts were added to the plates pre-coated with an oligonucleotide containing Nr2 consensus binding site and incubated for 1 h at RT. Then, the primary antibody was added to the wells and the plate was incubated for 1 h at RT. After washing, HRP-conjugated secondary antibody was added to the wells and the samples were incubated for 1 h at RT. Subsequently, each well was washed and a developing solution was added. When the solution color turned from medium to dark blue, a stop solution was added and the absorbance was measured at 450 nm.