Immunohistochemical analysis of PRAS40 in paraffin-embedded human breast tissue. Sample was incubated with polyclonal PRAS40 antibody (Product # PA5-86269).

Western blot was performed using Anti-PRAS40 Polyclonal Antibody (Product # PA5-86269) and a 40kDa band corresponding to Proline-rich AKT1 substrate 1 was observed across tissues tested. Whole cell extracts (30 µg lysate) of Mouse Liver (Lane 1), Rat Liver (Lane 2), Mouse Heart (Lane 3), Rat Heart (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 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).