Western blot was performed using CD166 (ALCAM) Polyclonal Antibody (Product # PA5-78755) and a 110 kDa band corresponding to ALCAM was observed across cell lines and mouse tissues. Whole cell extracts (30 µg lysate) of PC-3 (Lane 1), PC-3 treated with 1X Protein Transport Inhibitor (PTI) for 4hours (Lane 2), LNCaP (Lane 3), LNCaP treated with 1X Protein Transport Inhibitor (PTI) for 4hours (Lane 4), SH-SY5Y (Lane 5), Hep G2 (Lane 6), and tissue extracts of Mouse Liver (Lane 7), Mouse Kidney (Lane 8), Mouse Lung (Lane 9), Mouse Brain (Lane 10) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. 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 (1:1500) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).