Western blot was performed using Anti-OPG Mouse Monoclonal Antibody (Product # MA5-15715) and a 56 kDa band corresponding to OPG was observed in U-2 OS cell line treated with 1,3 - Cyclohexanebis (methylamine) mixture of isomers tested and not in SH-SY5Y which is reported to be negative. Whole cell extracts (30 µg lysate) of U-2 OS (Lane 1), U-2 OS treated with 1,3 - Cyclohexanebis (methylamine) mixture of isomers for 4 hours (Lane 2), SH-SY5Y (Lane 3) and U-2 OS treated with 1,3 - Cyclohexanebis (methylamine) mixture of isomers for 4 hours (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein 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 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).