Western blot analysis was performed on Whole cell extracts (30 µg lysate) of A549 (Lane 1), A549 (Condition Media) (Lane 2), PC-3 (Lane 3) and PC-3 (Condition Media) (Lane 4) and Tissue extracts of Mouse Colon (Lane 5) and Rat Stomach (Lane 6). The blots were probed with Anti-Anterior gradient homologue 2 (AGR2) Rabbit Polyclonal Antibody (Product # 720305, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 18 kDa band in whole cell extracts and 80 kDa band in condition media was observed corresponding to AGR2 protein across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), 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® 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).