VALIDATION IMAGES
4% Paraformaldehyde-fixed Hela (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (HSP90 beta) monoclonal Antibody, unconjugated (bsm-61128R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
25 ug total protein per lane of various lysates (see on figure) probed with HSP90 beta monoclonal antibody, unconjugated (bsm-61128R) 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 Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) 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 HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) 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 Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat
Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) 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 Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) 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 Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) 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 Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) 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 Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with HSP90 beta Monoclonal Antibody, Unconjugated(bsm-61128R) at 1:200 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
The Hela (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.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-HSP90 beta antibody (bsm-61128R): 1 μg/10^6 cells; Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.