Western blot analysis was performed on Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF7 (Lane 2), COS-7 (Lane 3), HEK-293 (Lane 4), C2C12 (Lane 5), Hep G2 (Lane 6), PANC-1 (Lane 7), PC-3 (Lane 8) and Caco-2 (Lane 9). The blots were probed with Anti-eIF4A1 Recombinant Rabbit Polyclonal Antibody (Product # 711505, 1-2 µg/mL). A 46 kDa band corresponding to eIF4A1 was observed across the cell lines tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).