Description
Principle of the Assay: Acetylcholine Assay Kit measures the acetylcholine present within serum, plasma, or tissue samples. The assay is based on the enzyme driven reaction that will detect acetylcholine via acetylcholinesterase enzyme and choline oxidase. First, acetylcholinesterase hydrolyzes acetylcholine into choline and acetic acid. Choline is then oxidized by choline oxidase to produce hydrogen peroxide. The hydrogen peroxide is then detected with a highly specific fluorescence probe. Horseradish peroxidase catalyzes the reaction between the probe and hydrogen peroxide, which bind in a 1:1 ratio. Samples are compared to a known concentration of acetylcholine standard within the 96-well microtiter plate format. Samples and standards are incubated for 45 minutes and then read with a standard 96-well fluorometric plate reader (Figure 1):