VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Caspase-3 Monoclonal Antibody, Unconjugated (bsm-55032M) at 1:200 overnight at 4°C, followed by conjugation to the bs-40296G-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 Caspase-3 Monoclonal Antibody, Unconjugated (bsm-55032M) at 1:200 overnight at 4°C, followed by conjugation to the bs-40296G-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; The section was incubated with Caspase-3 Monoclonal Antibody, Unconjugated (bsm-55032M) at 1:200 overnight at 4°C, followed by conjugation to the bs-40296G-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; The section was incubated with Caspase-3 Monoclonal Antibody, Unconjugated (bsm-55032M) at 1:200 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Jurkat (H) cells were treated with or without staurosporine treated (1uM) for 4 h, 25 μg total protein per lane of cell lysates (see on figure) probed with Caspase-3 monoclonal antibody, unconjugated (bsm-55032M) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
4% Paraformaldehyde-fixed Jurkat (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Caspase-3) monoclonal Antibody, unconjugated (bsm-55032M) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Mouse IgG antibody (green, bs-60296G-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.
The Jurkat (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):Mouse Anti-Caspase-3 antibody (bsm-55032M): 1 μg/10^6 cells; Isotype Control (orange): Mouse IgG (bs-0296P). Blank control (black): PBS. Acquisition of 20,000 events was performed.