Western blot analysis was performed on Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 (Condition media) (Lane 2), Caco-2 (Lane 3) and Caco-2 (Condition media) (Lane 4). The blots were probed with Anti-Vitronectin Rabbit Polyclonal Antibody (Product # 720229, 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 75 kDa band corresponding to Vitronectin was observed only in condition media 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).