Immunocytochemistry analysis of eIF4E in SK-Br-3 cells. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were incubated with eIF4E recombinant monoclonal antibody (Product # MA5-50717) with a dilution of 1:50 in 2% negative goat serum overnight at 4 ℃. Followed by secondary antibody Goat Anti-Rabbit IgG H&L (iFluor™ 488) at a dilution of 1:1,000. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.