| Abstract: | Specific enzymatic bands in disc gel electrophoresis are generally determined by either of two methods: (i) Gel is sliced and the enzymatic activity is assayed on each slice or (ii) gel is stained histochemically, if the product of the enzymatic reaction and the dye can form an insoluble precipitate, and the activity band is located on the gel by a color band. The former is laborious and often inaccurate in the calculation of electrophoretic mobility. The latter, often nonspecific, is not applicable when the enzymatic product cannot form an insoluble precipitate with the dye. Staining with tetrazolium salt has been widely employed for amine oxidase (1–6). However, this method has limitations: (i) Tetrazolium salt is nonspecific for amine oxidase and may show artifacts (6,7), and (ii) the use of tetrazolium salt is limited only to substrates containing indolamine such as tryptamine or serotonin (8). Other substrates, like benzylamine, the most active substrate for plasma amine oxidase, do not form a color band with tetrazolium salt.This communication reports a simple spectrophotometric method for the identification of the enzymatic activity band for amine oxidase on disc gel electrophoresis. Neither slice and assay nor staining is needed. This method may possibly also be used generally for other enzyme systems which have a specific absorption at ultraviolet or visible range. |
