Western blot was performed using Anti- ARF6 Polyclonal Antibody (Product # PA5-44191) and 20kDa band corresponding to ARF6 was observed across most of the cell lines and tissue extracts except Mouse Uterus which is reported negative for ARF6 expression. Differential expression of MCF 10A (lowest), MCF7 and MDA-MB-231 (highest) was also observed. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF 10A (Lane 2), MCF7 (Lane 3), MDA-MB-231 (Lane 4), HEK-293 (Lane 5), Hep G2 (Lane 6), COS-7 (Lane 7), Mouse Liver (Lane 8), Rat Liver (Lane 9) and Mouse Uterus (Lane 10) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blots were probed with the primary antibody (0.5 µg/mL) and detected by chemiluminescence Goat Anti-Rabbit IgG 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).