Anti-NF-kB p65 (phospho Ser529) Antibody (A306643)

Western blot analysis of extracts of various cell lines, using Anti-NF-kB p65 (phospho Ser529) Antibody (A306643) at 1:1,000 dilution. HeLa cells were treated by both TNF-alpha(20ng/ml) and Calyculin A (100nM) for 10 minutes. NIH/3T3 cells were treated by TNF-alpha(20ng/ml) for 30 minutes. The secondary antibody was Goat Anti-Rabbit IgG H&L Antibody (HRP) at 1:10,000 dilution. Lysates/proteins were present at 25µg per lane. The blocking buffer used was 3% BSA. Detection was with a ECL Basic Kit. Exposure time: 1s.

Western blot analysis of extracts of NIH/3T3 cells, using Anti-NF-kB p65 (phospho Ser529) Antibody (A306643) at 1:1,000 dilution or NF-kB p65 antibody (A16271). NIH/3T3 cells were treated by TNF-alpha (20 ng/ml) at 37°C for 30 minutes. The secondary antibody was Goat Anti-Rabbit IgG H&L Antibody (HRP) at 1:10,000 dilution. Lysates/proteins were present at 25µg per lane. The blocking buffer used was 3% BSA. Detection was with a ECL Basic Kit. Exposure time: 60s.

Immunohistochemistry analysis of paraffin-embedded rat lung using Anti-NF-kB p65 (phospho Ser529) Antibody (A306643) at a dilution of 1:100 (40x lens). Perform high pressure antigen retrieval with 10 mM citrate buffer pH 6.0 before commencing with IHC staining protocol.