Western blot analysis of extracts of NIH/3T3 and C6 cells, using Phospho-MTOR-S2448 antibody (144-50094) at 1:2000 dilution. NIH/3T3 cells were treated by Insulin (100nM) for 10 minutes after serum-starvation overnight. C6 cells were treated by IGF-1 (50 ng/ml) 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: 30S.
Immunofluorescence analysis of HeLa cells using Phospho-MTOR-S2448 antibody (144-50094) at dilution of 1:100. Blue: DAPI for nuclear staining.
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