Western blot was performed using Anti-APPL1 Rabbit Polyclonal Antibody (Product # PA5-76172) and a 79 kDa band corresponding to APPL1 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of SW480 (Lane 1), MCF7 (Lane 2), HeLa (Lane 3), HEK-293 (Lane 4), Mouse Brain (Lane 5), Rat Brain (Lane 6) and Mouse Spleen (Lane 7) were electrophoresed using 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 chemiluminescence with Goat anti-Rabbit IgG (H+L) 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). An uncharacterized around 45 kDa band (*) was observed in rat brain.