VALIDATION IMAGES
Paraformaldehyde-fixed, paraffin embedded (mouse thymus); Antigen retrieval by boiling in sodium EDTA buffer (Ph9.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Ki67) Monoclonal Antibody, Unconjugated (bsm-60738R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium EDTA buffer (Ph9.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Incubation with (Ki67) Monoclonal Antibody, Unconjugated (bsm-60738R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human Gastric Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated(bsm-60738R) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated(bsm-60738R) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated(bsm-60738R) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated(bsm-60738R) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated(bsm-60738R) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated(bsm-60738R) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023)and DAB (C-0010) staining.
4% Paraformaldehyde-fixed Hela(H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Ki-67) monoclonal Antibody, unconjugated (bsm-60738R) 1:200, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-0295G-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.
Primary Antibody (green line): Rabbit Anti-Iba1 antibody (bsm-60738R)
Dilution:0.5ug/Test;
Secondary Antibody(white blue line): Goat anti-rabbit IgG-FITC
Dilution: 0.5ug/Test.
Isotype control(orange line): Normal Rabbit IgG
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.