Western blot analysis of lysates from HeLa cells, using Phospho-NF-kB p65/RelA-S536 Rabbit mAb (AP1294) at1:2000 dilution.HeLa cells were treated by100ng/mL TNF-α + 50nM CA at 37℃ for 30 minutes after serum-starvation overnight
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.