Western blot analysis of extracts of HeLa cells, using Phospho-PKR/EIF2AK2-T446 antibody (TA375789) at 1:1000 dilution.HeLa cells were treated by Calyculin A (100 nM) at 37‚ÑÉ for 30 minutes after serum-starvation overnight.|Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution.|Lysates/proteins: 25ug per lane.|Blocking buffer: 3% nonfat dry milk in TBST.|Detection: ECL Basic Kit .|Exposure time: 90s.