Catalases and peroxidases histochemical detection; techniques suitable to discriminate these enzymes

Abstract

By using the benzidine reaction, on filter paper strips loaded with catalases, peroxidases, porphyrins, haemic iron compounds and iron salts, it was possible to establish 2 histochemical techniques able to detect and discriminate catalases and peroxidases. Spot test analytical studies show that only peroxidases oxidize benzidine in presence of a 0.0015 M H2O2 final concentration into the incubation medium. If a 0.0035 M H2O2 final concentration is used both peroxidases and haemic iron were able to oxidize benzidine. At a 0.01 M H2O2 final concentration the oxidative property of catalases become apparent and therefore at this H2O2 concentration either peroxidases or haemic iron, as well as catalases could be detected. By increasing the H2O2 concentration into the incubation medium, when a 4M concentration was chosen to detect histochemically catalases without any peroxidases interference. Using 0.0015 M and 4 M H2O2 final concentrations into the incubation medium it is possible to discriminate histochemically catalases and peroxidases. Several inhibitors of catalases and peroxidases were used as an attempt to try a specific inhibition of only one of these enzymes. It was demonstrated that the use of inhibitors does not help the histochemical discrimination between catalases and peroxidases.