Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1), MDCK (lane 2) and HEK-293 (Lane 3). The blots were probed with Anti-Claudin-16 Recombinant Rabbit Polyclonal Antibody (Product # 710220, 3-4 µg/mL) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). Isoforms of 26kDa and 34kDa bands corresponding to Claudin-16 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0341BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).