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In reaction mixture without enzyme solution, varying amounts of glutathione (0-300 l of stock solution giving 0-260 M final glutathione concentrations) were reacted with 150 l of monobromobimane solution (final concentration 50 M) in presence of phosphate buffer (3.0 ml) in cuvettes

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Fluorescence spectra were acquired using an FS5 spectrofluorometer (Edinburgh Instruments) equipped with an SC-40 plate reader and 96-well plates (Corning) using Fluoracle software