VALIDATION IMAGES
Formalin-fixed and paraffin embedded human colon carcinoma tissue labeled with Anti-phospho-Nrf2 (Ser40) Polyclonal Antibody (bs-2013R), Unconjugated 1:200 followed by conjugation to the secondary Cy5 antibody and DAPIstaining
Formalin-fixed and paraffin embedded human colon carcinoma tissue labeled with Anti-phospho-Nrf2 (Ser40) Polyclonal Antibody (bs-2013R), Unconjugated 1:200 followed by conjugation to the secondary Cy5 antibody
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Nrf2 (Ser40) Polyclonal Antibody, Unconjugated (bs-2013R) at 1:400 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Nrf2 (Ser40) Polyclonal Antibody, Unconjugated (bs-2013R) at 1:400 overnight at 4°C, DAB staining.
HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-Nrf2 (Ser40)) polyclonal Antibody, Unconjugated (bs-2013R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
25 ug total protein per lane of various lysates (see on figure) probed with phospho-Nrf2 (Ser40) polyclonal antibody, unconjugated (bs-2013R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
HeLa cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-Nrf2 (Ser40)) polyclonal Antibody, Unconjugated (bs-2013R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
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(bs-2013R)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).
Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Nrf2 (Ser40) Polyclonal Antibody, Unconjugated (bs-2013R) at 1:200 overnight at 4°C, 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; The section was incubated with phospho-Nrf2 (Ser40) Polyclonal Antibody, Unconjugated (bs-2013R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Nrf2 (Ser40) Polyclonal Antibody, Unconjugated (bs-2013R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.