Western blot was performed using AFP Polyclonal Antibody (Product # PA5-117771) and a 68 kDa band corresponding to AFP was observed only in Hep G2 cell line. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), MCF7 (Lane 2), A549 (Lane 3), HeLa (Lane 4), SK-O-V3 (Lane 5), A-431 (Lane 6), SH-SY5Y (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556).