Western blot was performed using Anti-AMPK gamma-1 Polyclonal Antibody (Product # PA5-27471) and a 36 kDa band corresponding to AMPK gamma-1 was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of PC-3 (Lane 1) and T-47D (Lane 2) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0302BOX). 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 chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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). Additional bands at ~65kDa and !51 kDa were observed which corresponds to AMPK gamma-2 and AMPK gamma-3 respectively.