Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), COLO 205 (Lane 3), SH-SY5Y (Lane 4), Mouse Brain (Lane 5) and Rat Brain (Lane 6). The blots were probed with Anti-RhoB Recombinant Rabbit Polyclonal Antibody (Product # 711274, 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 22 kDa band corresponding to RhoB was observed across cell lines 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).