Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse,Rat

IgG

Human,Mouse,Rat

1. sample: A549 cells ((blue) Ice-cold 70% ethanol was added and the cells were incubated for overnight at 4°C. and then resuspended in ice-cold 90% methanol for 30 min on ice.)
Primary Antibody: Rabbit Anti-PCNA antibody (TMAB-01342) (green)
Isotype Control Antibody: Rabbit IgG (orange)
Secondary Antibody: f (ab/)2 fragment goat anti-rabbit IgG-FITC (white blue)
2. Blank control (blue line): U251 (blue).
Primary Antibody (green line): Rabbit Anti-PCNA antibody (TMAB-01342)
Dilution: 3 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. 25 μg total protein per Lane of various lysates probed with PCNA polyclonal antibody, unconjugated (TMAB-01342) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.
4. Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Human Liver Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded Mouse Testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
15. Paraformaldehyde-fixed, paraffin embedded Rat Testis; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
16. Paraformaldehyde-fixed, paraffin embedded Mouse Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
17. Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
18. Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
19. Paraformaldehyde-fixed, paraffin embedded Rat Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
20. Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Polyclonal Antibody, Unconjugated (TMAB-01342) at 1:500 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:200-800; IHC-Fr: 1:200-800; IF: 1:200-800; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human PCNA

Human

5111

Cell division,Tumor antigens,Markers,DNA / RNA binding,DNA Synthesis,Methylated DNA Binding,PCNA,Replication