Differential identification of mouse granulocyte (CFU-g) and macrophage (CFU-m) precursors in plasma clots.
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.
Iizuka
Y
Y
Murphy
M J
MJ
D000818:Animals; D001560:Benzidines / Q000378:metabolism; D002396:Catechols / Q000378:metabolism; D006098:Granulocytes / Q000201:enzymology / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000145:classification* / Q000201:enzymology; D008264:Macrophages / Q000201:enzymology / Q000502:physiology; D008297:Male; D051379:Mice; D009283:Naphthol AS D Esterase / Q000378:metabolism; D010544:Peroxidases / Q000378:metabolism; D010655:Phenylenediamines / Q000378:metabolism; D010949:Plasma; D013194:Staining and Labeling
D001560:Benzidines; D002396:Catechols; D010655:Phenylenediamines; C029876:benzidine; D010544:Peroxidases; D009283:Naphthol AS D Esterase; C034221:catechol; C029728:4-phenylenediamine