The classical role of ACHE is to terminate cholinergic neurotransmission by hydrolysis of acetylcholine (ACH). ACHE is thought to be involved in the pathology of Alzheimer's disease (AD) by accelerating the assembly of A beta peptides into fibrillar species through forming complexes with A beta via the peripheral anionic site on ACHE. ACHE inhibitors have been used to delay symptoms of AD patients by virtue of their ability to enhance ACH availability, as well as reduce amyloidogenesis and subsequent neurotoxicity. Its involvement in the cholinergic anti-inflammatory pathway connects ACHE with a possible marker of low-grade systemic inflammation in obesity, hypertension, coronary heart disease, and AD. The activity of recombinant mouse ACHE was measured by its ability to cleave Acetylthiocholine in the assay buffer 0.1 M sodium phosphate, 0.05% (w/v) Brij-35, pH 7.5. 50 μL of various concentrations of rmACHE (diluted by Assay Buffer) was added into the 96-well clear plate and the reaction was started by the addition of 50 µl substrate mixture of 200 uM acetylthiocholine and 100 uM DTNB. The final well serves as a negative control with no rmACHE, replaced with 50μl assay buffer and 50 ul substrate mixture. Read plate in kinetic mode for 5 minutes at an absorbance of 405 nm. The specific activity of recombinant mouse ACHE is >2200 nmol/min/µg.