Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Nrf2(S40) (7G4) Monoclonal Antibody(bsm-52179R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-Nrf2 (Ser40)) monoclonal Antibody, Unconjugated (bsm-52179R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded Human Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated(bsm-52179R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated(bsm-52179R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.

25 ug total protein per lane of various lysates (see on figure) probed with phospho-Nrf2 (Ser40) monoclonal antibody, unconjugated (bsm-52179R) at 1:2000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated(bsm-52179R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Skin Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated(bsm-52179R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated(bsm-52179R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated (bsm-52179R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.