Left: Human topo I (S 35 Met-labeled) control. Right: Sumoylated human topo I (S 35 Met-labeled) (7.5 µg/ml E1, 50 µg/ml E2, 50 µg/ml sumo1, 37ºC 30min).
Western Blot: Extracts prepared from CEF cells not transfected (-) or transfected (+) with wild-type Src were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 1.0 µ g/mL anti-Src pan antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected using the Tropix WesternStar method.
Extracts prepared from CEF cells not transfected (-) or transfected (+) with wild-type Src were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 1.0 µg/mL anti-Src pan antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected using the Tropix WesternStar method.
