Western blot was performed using Anti-AFP Antibody (Product # PA5-78734) and a 68 kDa band corresponding to AFP was observed in Hep G2 cells and Mouse Placenta but not in MDA-MB-231 cells, Mouse Lung and Mouse Liver which are reported to be low to negative for AFP expression. Additional faint uncharacterized band (*) was observed across the panel. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with 1X protein transport inhibitor (Product # 00-4980-03) for 4 hours (Lane 2), MDA-MB-231 (Lane 3) and MDA-MB-231 treated with 1X protein transport inhibitor for 4 hours (Product # 00-4980-03) (Lane 4), tissue extracts of Mouse Placenta (Lane 5), Mouse Liver (Lane 6) and Mouse Lung (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).