Western blot was performed using Anti-RAP1A Monoclonal Antibody (5F8) (Product # MA5-17164) and 20 kDa band corresponding to RAP1A was observed across the cell lines tested except C2C12 in which the expression was observed upon myoblast differentiation. Whole cell lysates (30ug lysate) of C2C12 (Lane 1), C2C12 differentiated to Myoblasts (Lane2), HT-29 (Lane 3), MDA-MB-231 (Lane 4), MCF 10A (Lane 5), A-431 (Lane 6), HeLa (Lane 7) and A-375 (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).