Silveira S R, Hadler W A
Acta Histochem. 1978;63(1):1-10. doi: 10.1016/S0065-1281(78)80002-6.
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.
通过使用联苯胺反应,在载有过氧化氢酶、过氧化物酶、卟啉、血红素铁化合物和铁盐的滤纸条上,有可能建立两种能够检测和区分过氧化氢酶和过氧化物酶的组织化学技术。点滴试验分析研究表明,在孵育介质中最终过氧化氢浓度为0.0015M时,只有过氧化物酶能将联苯胺氧化。如果使用最终过氧化氢浓度为0.0035M,则过氧化物酶和血红素铁都能氧化联苯胺。在最终过氧化氢浓度为0.01M时,过氧化氢酶的氧化特性变得明显,因此在这个过氧化氢浓度下,可以检测到过氧化物酶或血红素铁以及过氧化氢酶。通过提高孵育介质中的过氧化氢浓度,当选择4M浓度进行组织化学检测过氧化氢酶时,不会有任何过氧化物酶的干扰。在孵育介质中使用最终浓度为0.0015M和4M的过氧化氢,可以在组织化学上区分过氧化氢酶和过氧化物酶。使用了几种过氧化氢酶和过氧化物酶抑制剂,试图仅对其中一种酶进行特异性抑制。结果表明,使用抑制剂无助于在组织化学上区分过氧化氢酶和过氧化物酶。