Choline acetyltransferase (ChAT) mediates the synthesis of the neurotransmitter acetylcholine from acetyl-CoA and choline. In addition to its critical role in neurosignaling, deficiencies in ChAT are linked to congenital myasthenic syndromes, Alzheimer’s disease, and multiple sclerosis. α-NETA is a stable, non-competitive, and slowly reversible inhibitor of ChAT (IC50 = 9 μM). This naturally fluorescent compound is a poor inhibitor of cholinesterases and carnitine acetyltransferase. α-NETA is commonly used in cells in culture.