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 ((treated with MG-132(10 μM,10h)) ) ; 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.
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.
The HeLa (treated with MG-132(10 μM,10 h)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-Nrf2 (Ser40) antibody (bs-2013R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.