Western blot analysis was performed on whole cell extracts of A-431 (Lane 1), HEK-293 (Lane 2), HeLa (Lane 3), Hep G2 (Lane 4) and U-87 MG (Lane 5). The blot was probed with Anti-4EBP1 antibody (Product # AHO1382, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 17 kDa band corresponding to 4EBP1 was observed across the cell lines tested.