VALIDATION IMAGES
4% Paraformaldehyde-fixed Hela (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Ki-67) monoclonal Antibody, unconjugated (bsm-0061R) 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 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):Mouse Anti-Ki-67 antibody (bsm-258161M): 1 μg/10^6 cells; Isotype Control (orange): Mouse IgG (bs-0296P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
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 Ki-67 Monoclonal Antibody, Unconjugated (bsm-25816M) at 1:1000 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Serous ovarian cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Ki-67 Monoclonal Antibody, Unconjugated (bsm-25816M) at 1:1000 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Ki-67 Monoclonal Antibody, Unconjugated (bsm-25816M) at 1:1000 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 Ki-67 Monoclonal Antibody, Unconjugated (bsm-25816M) at 1:1000 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
(Negative control) Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Ki-67 Monoclonal Antibody, Unconjugated (bsm-25816M) at 1:1000 overnight at 4°C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.