Western blot was performed using Anti-ASCL1 Polyclonal Antibody (Product # PA5-77868) and 32 kDa band was observed corresponding to ASCL1 was observed across the cell lines tested along with an uncharacterized band (*) in Mouse pup brain. Modified Whole cell extracts (30ug lysate) (1% SDS) of Neural Stem cells (Lane 1), SH-SY5Y (Lane 2), U-87 MG (Lane 3), IMR-32 (Lane 4), Neuro-2A (Lane 5), tissue extracts of Mouse pup brain (Lane 6) and Rat pup brain (Lane 7) were electrophoresed using Novex® 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 (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)..