Western blot analysis of MDM2 in extracts of NIH/3T3 cells. Samples were incubated with MDM2 Polyclonal antibody (Product # PA5-87423) using a dilution of 1:1,000, followed by HRP Goat Anti-Rabbit IgG (H+L) secondary antibody at a dilution of 1:10,000. NIH/3T3 cells were treated by MG132 (50 μM) at 37°C for 90 minutes. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 90s.