Immunohistochemical analysis of Phospho-eIF4E (Ser209) in paraffin embedded human breast cancer using a Phospho-eIF4E (Ser209) monoclonal antibody (Product # MA5-33134) at a dilution of 1:100. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Western Blot analysis of Phospho-eIF4E (Ser209) using a Phospho-eIF4E (Ser209) Monoclonal antibody (Product # MA5-33134) at a concentration of 1.2 µg/mL. Positive WB detected in: 293 whole cell lysate,HepG2 whole cell lysate(treated with Calyculin A or not). All lanes: Phospho-EIF4E antibody at 1.2µg/ml. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 25 kDa.