Western blot analysis was performed on Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), U-87 MG (Lane 2) and HeLa (Lane 3). The blots were probed with Anti-Spatacsin Recombinant Rabbit Polyclonal Antibody (Product # 711811, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A 278 kDa band corresponding to Spatacsin was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and HiMark™ Pre-Stained Protein Standard (Product # LC5699). Resolved proteins were then transferred onto a nitrocellulose membrane by wet transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).