Anti-PCNA Antibody (3D152), Nuclear Loading Control is a Rabbit antibody targeting PCNA. Anti-PCNA Antibody (3D152), Nuclear Loading Control can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

3D152

Human,Mouse,Rat

1. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-PCNA antibody (TMAB-10076)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
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℃. 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.
2. Paraformaldehyde-fixed, paraffin embedded Mouse Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded Rat Heart; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded Human Liver Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded Human Duodenum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA (Nuclear Loading Control) Monoclonal Antibody, Unconjugated (TMAB-10076) at 1: 200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Monoclonal Antibody, Unconjugated (TMAB-10076) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Monoclonal Antibody, Unconjugated (TMAB-10076) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded Rat Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Monoclonal Antibody, Unconjugated (TMAB-10076) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
15. Paraformaldehyde-fixed, paraffin embedded Mouse Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Monoclonal Antibody, Unconjugated (TMAB-10076) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
16. Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA Monoclonal Antibody, Unconjugated (TMAB-10076) at 1:800 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.
17. 25 μg total protein per lane of various lysates (see on figure) probed with PCNA monoclonal antibody, unconjugated (TMAB-10076) at 1:10000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r. T. for 60 min.

WB: 1:1000-20000; IHC-P: 1:200-1000; IHC-Fr: 1:200-1000; ICC/IF: 1:100-500; IF: 1:200-1000; FCM: 1μg/Test

Monoclonal

A synthesized peptide: human PCNA

Human

PCNA

Proliferating cell nuclear antigen

Auxiliary protein of DNA polymerase delta and epsilon, is involved in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leading strand (PubMed:35585232).Induces a robust stimulatory effect on the 3'-5' exonuclease and 3'-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. Has to be loaded onto DNA in order to be able to stimulate APEX2. Plays a key role in DNA damage response (DDR) by being conveniently positioned at the replication fork to coordinate DNA replication with DNA repair and DNA damage tolerance pathways (PubMed:24939902).Acts as a loading platform to recruit DDR proteins that allow completion of DNA replication after DNA damage and promote postreplication repair: Monoubiquitinated PCNA leads to recruitment of translesion (TLS) polymerases, while 'Lys-63'-linked polyubiquitination of PCNA is involved in error-free pathway and employs recombination mechanisms to synthesize across the lesion (PubMed:24695737).