Iizuka Y, Murphy M J
Int J Cell Cloning. 1985 May;3(3):176-84. doi: 10.1002/stem.5530030306.
Because benzidine and its derivatives have possible carcinogenic activity, a safe method is needed to demonstrate endogenous peroxidase activity. Colonies derived from mouse bone marrow cells in plasma clot culture were classified as granulocyte (CFU-g) or macrophage (CFU-m) precursors by peroxidase and naphthol AS acetate (NASA) esterase staining, respectively. Endogenous peroxidase activity was measured using benzidine or p-phenylenediazine-pyrocatechol (PPD-PC). The effectiveness of peroxidase staining with both reagents was evaluated under several conditions, and the enzyme property was confirmed by inactivation with a variety of inhibitors. The level of peroxidase activity did not differ significantly between PPD-PC and benzidine. Colony number and number of cultured cells were strongly correlated (P greater than 0.983). We conclude that PPD-PC safely demonstrates peroxidase activity in cultured cells and is as accurate, reliable, and efficient as benzidine.
由于联苯胺及其衍生物具有潜在致癌活性，因此需要一种安全的方法来检测内源性过氧化物酶活性。通过过氧化物酶和萘酚AS醋酸酯（NASA）酯酶染色，分别将血浆凝块培养中源自小鼠骨髓细胞的集落分类为粒细胞（CFU-g）或巨噬细胞（CFU-m）前体。使用联苯胺或对苯二胺-邻苯二酚（PPD-PC）测量内源性过氧化物酶活性。在几种条件下评估了两种试剂进行过氧化物酶染色的效果，并通过用多种抑制剂使其失活来确认酶的性质。PPD-PC和联苯胺之间的过氧化物酶活性水平没有显著差异。集落数与培养细胞数密切相关（P大于0.983）。我们得出结论，PPD-PC能安全地检测培养细胞中的过氧化物酶活性，并且与联苯胺一样准确、可靠和高效。
