A fluorometric assay for acetylcholine with picomole sensitivity.
A sensitive fluorogenic assay for acetylcholine is described. The assay is based upon the reactions: (1) hydrolysis of acetylcholine to choline and acetate, catalyzed by acetylcholinesterase. (2) Oxidation of choline to betaine and hydrogen peroxide by choline oxidase. (3) Oxidation of p-hydroxyphenylacetic acid by hydrogen peroxide to a fluorescent product, catalyzed by peroxidase. With a sensitivity comparable to chemiluminescence procedures, the assay should find application to those situations requiring the persistence of a fluorescence signal or the necessity of assaying small volumes.
Department of Biochemistry, Molecular and Cell Biology, Northwestern University, Evanston, IL 60208.
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MacDonald
D000109:Acetylcholine / Q000032:analysis*; D000818:Animals; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000379:methods*
D000109:Acetylcholine