Western blot was performed using Anti-CLDN1 Polyclonal Antibody (Product # PA1-37465) and a 22 kDa band corresponding to CLDN1 was observed across cell lines tested and increased upon EGF, LiCl treatment and IL-beta treatment. Membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), MDCK (Lane 2), A-431 (Lane 3), HEK-293 (Lane 4), Caco-2 treated with EGF (100 ng/mL) and LiCl (50mM) simultaneously for 24 Hours (Lane 5), Caco-2 treated with IL-beta (10 ng/mL for 48 Hours) (Lane 6), Caco-2 (Lane 7), tissue extracts (30 µg lysate) of Mouse Kidney (Lane 8), Mouse Liver (Lane 9), Mouse Colon (Lane 10) and Mouse Lung (Lane 11) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 (1:1,000 dilution) 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).