Western blot analysis of extracts of various cell lines, using Phospho-S6 Ribosomal Protein -S235/236 antibody at 1:1000 dilution.HeLa cells NIH/3T3 cells and PC-12 cells were treated by 10% FBS at 37℃ for 30 minutes after serum-starvation overnight.NIH/3T3 cells were treated by PDGF at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP Goat Anti-Rabbit IgG at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit .
Exposure time: 1s.
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