Western blot analysis of extracts of various cell lines, using Phospho-ERK1-T202/Y204 + ERK2-T185/Y187 antibody (TA378294) at 1:1000 dilution.Jurkat cells and NIH/3T3 cells and C6 cells were treated by PMA/TPA (200 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 Enhanced Kit .|Exposure time: 180s.