H. Maeda, H. Matsuno, M. Ushida
May 6, 2005
Citations
3
Influential Citations
333
Citations
Quality indicators
Journal
Angewandte Chemie
Abstract
Since its first description in 1959, 5,5’-dithio-bis(2-nitrobenzoic acid) (DTNB or Ellman s reagent) has been used widely for the quantification of thiols, especially for assays of various enzymes including acetyland butyrylcholinesterase (AChE and BChE, respectively) in which substrates release thiols through enzymatic reactions. 4,4’-Bipyridyl disulfide and 5-(2-aminoethyl)dithio-2-nitrobenzoic acid were also shown to be useful for colorimetric analyses of thiols. However, fluorescent or chemiluminescent thiol probes would be more practical than these chromogenic probes for enzyme assays, especially for high-throughput programs to screen enzyme inhibitors as new drugs. Recently, the use of a dioxetane-disulfide derivative as a chemiluminescent probe in place of DTNB led to an improved sensitivity of the AChE assay with acetylthiocholine (ASCh) by a factor of ten. However, the assay required a rather tedious procedure in