Western blot was performed using Anti-ITGA4 Polyclonal Antibody (Product # PA5-20595) and 160, 150 kDa bands corresponding to ITGA4 were observed. Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), HEL 92.1.7 (Lane 3), THP-1 (Lane 4), Hep G2 (Lane 5), MCF7 (Lane 6) and K-562 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Hep G2, MCF7 and K-562 have been reported to express ITGA4 at very low levels.