Western blot was performed using Anti-NEFL Polyclonal Antibody (Product # PA3-16719) and a 68 kDa band corresponding to NEFL was observed in PC-12 differentiated to neurons, Mouse Cerebellum, Mouse Brain and Rat Brain but not in PC-12, Mouse Heart and Mouse Lung along with few uncharacterized bands (*). Membrane enriched extracts (30 µg lysate) of PC-12 (Lane 1), PC-12 differentiated to neurons (Lane 2), Mouse Cerebellum (Lane 3), Mouse Brain (Lane 4), Rat Brain (Lane 5), Mouse Heart (Lane 6) and Mouse Lung (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:20,000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).* - There were few uncharacterized bands observed in PC-12 differentiated to neurons, Mouse Cerebellum and Mouse Brain.