In research paper generally researchers had been used Lester's method to measure ascorbate peroxidase activity and peroxidase activity. But I did not reach Lester's article related to these issue. So anybody can tell me alternative methods for measurement.
Hi, mehtap, there are many method for measure of APX and peroxidase activity. You can find them in couple of the articles. There are two basical way: measure total activity and separate isoform. In many case information about activity of each isoform is even more importnat to comapre with total activities. because in many cases activity of one isoform is dowm regulate, but the othjer is up-reguklate. In this case you need to prepare native PAAG and measure activity after isoform separation.
A common procedure is the determination of Peroxidase enzymatic activity using Pyrogallol as the substrate:
H2O2 + Pyrogallol --> 2H2O + purpurgallin
Reaction can be monitored by spectrophotometric assay measuring absorbance value at 420 nm. One unit of peroxidase will form 1.0 milligram of purpurogallin from pyrogallol in 20 seconds at pH 6.0 at 20 °C.
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