Western blot analysis of various lysates using Phospho-STAT1-Y701 Rabbit mAb (AP0054) at 1:1000 dilution incubated overnight at 4℃. HeLa cells, NIH/3T3 cells and PC-12 cells were treated with TNF-α (100 ng/mL) at 37℃ for 30 minutes.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 90s.