Western blot was performed using Anti-ChAT Monoclonal Antibody (CL3173) (Product # MA5-31383) and a ~70 kDa band corresponding to acetyl CoA: choline O-acetyltransferase was observed in Mouse and Rat Brain lysates but not in Mouse and Rat Kidney lysates. Whole cell extracts (30 µg lysate) of Mouse Brain (Lane 1), Rat Brain (Lane 2), Mouse Kidney (Lane 3) and Rat Kidney (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. 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 (1:500 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Mouse Light chain band (*) was observed at ~25 kDa in Mouse Brain and Kidney lysates and an uncharacterized band (*) was observed in Rat Kidney lysates ~90 kDa.