[IF=3.881] Zubeyir Elmazoglu. et al. Cannabinoid-profiled agents improve cell survival via reduction of oxidative stress and inflammation, and Nrf2 activation in a toxic model combining hyperglycemia+Aβ1-42 peptide in rat hippocampal neurons. Neurochem Int. 2020 Nov;140:104817  WB ;  Rat.  

PubMed:32781098

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°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

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°C, 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°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

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°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; Antibody incubation with phospho-Nrf2 (Ser40) Monoclonal Antibody, Unconjugated(bsm-52179R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.

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°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.

MCF7 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)) monoclonal Antibody, Unconjugated (bsm-52179R) 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.

HUVEC 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)) monoclonal Antibody, Unconjugated (bsm-52179R) 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.

Blank control:THP-1. Primary Antibody (green line): Rabbit Anti-phospho-Nrf2 (Ser40) antibody (bsm-52179R) Dilution: 1ug/Test; Secondary Antibody : Goat anti-rabbit IgG-FITC Dilution: 0.5ug/Test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.