Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (Lane 1), Hep G2 (Lane 2), K562 (Lane 3), HEK-293 (Lane 4), NTERA-2 (Lane 5), Cos-7 (Lane 6), HeLa (Lane 7), Jurkat (Lane 8), MCF7 (Lane 9), PC-3 (Lane 10). The blots were probed with Anti-RNF20 Recombinant Rabbit Polyclonal Antibody (Product # 710911, 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 120 kDa band corresponding to RNF20 was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10% Bis-Tris gel (Product # NP0301BOX), 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 PierceTM Power Blotter System (Product # 22834). 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).