ab76256 staining eIF4E (phospho S209) in serum starved HEK193 cells treated with CGP 57380 (ab120365), by ICC/IF. Decrease in eIF4E (phospho S209) expression correlates with increased concentration of CGP 57380, as described in literature.The cells were incubated at 37°C for 1h in media containing different concentrations of ab120365 (CGP 57380) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab76256 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.