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收藏Western blot analysis of extracts of various cell lines, using Phospho-ERK1-T202/Y204 + ERK2-T185/Y187 antibody (144-50472) at 1:5000 dilution or ERK1/2 antibody (144-613146).293T cells and C6 cells were treated by ATP(5 mM) at 30? for 1 hour. NIH/3T3 cells were treated by PMA/TPA (200 nM) at 37? 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% BSA.
Detection: ECL Basic Kit.
Exposure time: 10s.
Immunohistochemistry of paraffin-embedded rat lung using Phospho-ERK1-T202-Y204 + ERK2-T185-Y187 antibody (144-50472) at dilution of 1:100 (40x lens).