Immunocytochemistry-Immunofluorescence analysis of GAP43 was performed in DIV9 rat E18 primary cortical neurons fixed in 4% paraformaldehyde at RT for 15 min. Green: GAP43 Polyclonal Antibody (Product # PA5-27448) diluted at 1:500. Red: beta Tubulin 3/ Tuj1, stained by beta Tubulin 3/ Tuj1 antibody. Blue: Fluoroshield with DAPI.

Western blot was performed using Anti- GAP43 Polyclonal Antibody (Product # PA5-27448 ) and bands around 38-48 kDa corresponding to GAP43 was observed in Mouse and Rat Brain but not in other tissues tested, which are reported to be negative for GAP43 expression. Additional faint uncharacterized band (*) was observed in Rat Brain. Tissues extracts (20 µg lysate) of Mouse Lung(Lane 1), Rat Lung (Lane 2), Mouse Kidney (Lane 3), Rat Kidney (Lane 4), Mouse Brain (Lane 5) and Rat Brain (Lane 6) were electrophoresed using NuPAGE® 4-12 % Bis-Tris 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 (Heavy Chain), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/mL, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).