Western blot analysis of extracts of various cell lines, using Phospho-CHUK-S176 at 1:1000 dilution.HeLa cells were treated by TNF-alpha (20ng/ml) at 37℃ for 30 minutes.C6 cells were treated by TNF-alpha (20ng/ml) at 37℃ for 30 minutes. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L)at 1:10,000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% BSA. Detection: ECL Basic Kit Exposure time: 30s.