Western blot analysis was performed on Whole cell extracts (30 µg lysate) of 3T3L1 (Lane 1), LNCap (Lane 2) and tissue extracts of Rat Adipose (Lane 3) and Mouse skin (Lane 4). The blots were probed with Anti-PRDM16/MEL1 Rabbit Polyclonal Antibody (Product # 720206, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Two isoforms of 140 and 120 kDa corresponding to PRDM16/MEL1 was observed. 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).