Western blot was performed using Anti-CD13 Polyclonal Antibody (Product # PA5-109314) and a ~150 kDa band corresponding to ANPEP was observed across across cell lines tested. Membrane enriched extracts (30 µg lysate) of A-431 (Lane 1), THP-1 (Lane 2), PC-3 (Lane 3), Hep G2 (Lane 4), HL-60 (Lane 5), HUVEC (Lane 6) and Mouse Kidney (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).