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False positiveness in a choline acetyltransferase assay caused by nonionic reducing reagents
Author(s) -
Kobayashi Y.,
de Vellis J.
Publication year - 1985
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490130405
Subject(s) - dithiothreitol , reagent , choline acetyltransferase , chemistry , acetyltransferase , choline , acetylcholine , reducing agent , enzyme , chromatography , biochemistry , pharmacology , organic chemistry , medicine , acetylation , gene
Dithiothreitol and 2‐mercaptoethanol have been frequently used as protective agents of choline acetyltransferase (EC 2. 3. 1. 6) during its purification. We found that those reagents reacted with acetyl‐CoA to form products that were co‐extracted with acetylcholine into organic scintillation liquid to be erroneously detected as high enzyme activity. An anionic reducing reagent, such as sodium thioglycolate, does not cause the above problem and can be an alternative protective agent.