Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), MDA-MB-231 (Lane 2), T-47D (Lane 3), U-87 MG (Lane 4), Hep G2 (Lane 5) and tissue extracts of Mouse Kidney (Lane 6), Rat Kidney (Lane 7), Mouse Testis (Lane 8) and Rat Testis (Lane 9). The blots were probed with Anti-ELF5 Rabbit Polyclonal Antibody (Product # 720380, 1-2 µg/mL). An isoform of 31 kDa was observed in all cell lines tested and 16 kDa was observed in all tissues tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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 iBlot® Dry Blotting System (Product # IB21001). 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).