Immunohistochemistry analysis of NTCP on paraffin-embedded human liver cancer tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with NTCP polyclonal antibody (Product# PA5-80001) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Western Blot was performed using Anti-NTCP Polyclonal Antibody (Product # PA5-80001) and 40-50 kDa bands corresponding to Sodium/bile acid cotransporter and glycosylated form of NTCP were observed in Mouse Liver and Rat Liver but absent in Mouse Brain and Rat Brain. Tissue extracts (30 µg lysate) of Mouse Liver (Lane 1), Rat Liver (Lane 2), Mouse Brain (Lane 3) and Rat Brain (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (at a concentration of 0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).