Antibody Type: Recombinant Rabbit Monoclonal

Application: IHC-P,IHC-Fr,ICC/IF,IF,FCM

Reactivity: Human,Mouse,Rat

IgG

3H14

Human,Mouse,Rat

1. Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium EDTA buffer (Ph9.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Incubation with (Ki67) Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (mouse thymus); Antigen retrieval by boiling in sodium EDTA buffer (Ph9.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Incubation with (Ki67) Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-Ki67 antibody (TMAB-01142)
Dilution: 0.5 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C, 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.
4. 4% Paraformaldehyde-fixed Hela (H) cell; Triton X-100 at RT for 20 min; Antibody incubation with (Ki-67) monoclonal Antibody, unconjugated (TMAB-01142) 1:200, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green) at 37°C for 90 min, DAPI (blue) was used to stain the cell nucleus. PBS instead of the primary antibody was used as the blank control.
5. (Negative control) Paraformaldehyde-fixed, paraffin embedded Mouse Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
6. (Negative control) Paraformaldehyde-fixed, paraffin embedded Rat Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
7. (Negative control) Paraformaldehyde-fixed, paraffin embedded Human Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
8. 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 (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
9. 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 (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
10. 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 (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
11. 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 (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
12. 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 (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
13. 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 (TMAB-01142) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
15. Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.
16. Paraformaldehyde-fixed, paraffin embedded Human Esophageal Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Ki67 Monoclonal Antibody, Unconjugated (TMAB-01142) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (green), DAPI (blue) was used to stain the cell nucleus.

IHC-P,IHC-Fr,ICC/IF,IF,FCM

IHC-P: 1:200-800; IHC-Fr: 1:200-800; ICC/IF: 1:200; IF: 1:200-800; FCM: 0.5ug/Test

Monoclonal

KLH conjugated synthetic peptide: human Ki67

Human

4288

Cell division,Tumor biomarkers,Markers,Soma marker,Neurogenesis,Replication