The appearance of quinoneimine dye formed when coupled with 4-aminoantipyrine and phenol is measured at 500nm by spectrophotometry.
One unit causes the formation of one micromole of hydrogen peroxide (half a micromole of quinoneimine dye) per minute under the conditions described below.
Concentration in assay mixture
At the same time, measure the blank rate (ΔOD blank) using the same method as the test except that the enzyme diluent (G) is added instead of the enzyme solution.
* Dissolve the enzyme preparation in ice-cold enzyme diluent (G), and dilute to 0.08－0.22U/ml with the same buffer, immediately before assay.
Activity can be calculated by using the following formula：
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