PCNA Antibody

Staining PCNA in human breast carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

All lanes: Anti-PCNA Antibody at1: 3000 dilution. Lane 1: Hela whole cell lysate. Lane 2: A431 whole cell lysate. Lane 3: HepG2 whole cell lysate. Lane 4: human spleen lysate. Lane 5: mouse testis lysate. Lane 6: C2C12 whole cell lysate. Lane 7: rat liver lysate. Lane 8: C6 whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 36 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

All lanes: Anti-PCNA Antibody at 1:5000 dilution. Lane 1: Hela whole cell lysate. Lane 2: A431 whole cell lysate. Lane 3: A549 whole cell lysate. Lane 4: HepG2 whole cell lysate. Lane 5: NIH/3T3 whole cell lysate. Lane 6: C2C12 whole cell lysate. Lane 7: PC-12 whole cell lysate. Lane 8: C6 whole cell lysate. Lysates/proteins at 20 µg per lane. Secondary Goat Anti-mouse IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size: 36 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Overlay histogram showing Hela cells stained (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37°C. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at 37°C. Isotype control antibody (blue line) was mouse IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of >10000 events was performed.