Western blot was performed using Anti-NUMB Monoclonal Antibody (Product # MA5-32568) on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), PC-3 (Lane 2), DU 145 (Lane 3), SH-SY5Y (Lane 4), HeLa (Lane 5), Mouse lung (Lane 6) and Mouse Kidney (Lane 7) and 2 numb isoforms of 65 kDa and 55 kDa were observed. 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 (1:2000 dilution) and detected by Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).