Western blot was performed using Glypican 3 Polyclonal Antibody (Product # PA5-47256) and a 66 kDa band corresponding to GPC3 was observed only in Hep G2. Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3) and T-47D (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 (1:500 dilution) and detected by chemiluminescence with Rabbit anti-Sheep IgG (H+L), Secondary Antibody, HRP (Product # 61-8620, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).