Immunohistochemistry analysis of RNF2 in paraffin-embedded human breast cancer tissue. Samples were incubated with RNF2 polyclonal antibody (Product # PA5-76758).

Western blot was performed using Anti-RNF2 Polyclonal Antibody (Product # PA5-76758) and a ~38kDa band corresponding to RNF2 was observed across all the cell lines tested except in Raji, which is a low expressing cell line for the same along with an uncharacterized band at ~60kDa across the panel. Modified whole cell extracts (1% SDS) (30 µg lysate) of K-562 (Lane 1), HCT 116 (Lane 2), Jurkat (Lane 3), MOLT-4 (Lane 4), Raji (Lane 5), NIH/3T3 (Lane 6) and PC-12 (Lane 7)were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 (1:1200 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain), Superclonal™ Recombinant Secondary Antibody, HRP (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).