Western blot analysis of extracts of NIH/3T3 cells, using Phospho-CDK4-T172 pAb at 1:1000 dilution or CDK4 antibody .NIH/3T3 cells were treated by Hydroxyurea at 37℃ for 20 hours.
Secondary antibody: HRP Goat Anti-Rabbit IgG at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit .
Exposure time: 60s.
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