Abstract
A cyanide-insensitive b-type cytochrome (cytochrome b 559) is involved in the respiratory burst of phagocytes. Spectroscopic measurement of its content in polymorphonuclear leukocytes or macrophages is complicated by signals arising from contaminating Hb or mitochondrial cytochromes upon reduction by dithionite. These interferences can be overcome by reducing the sample with dithionite, aerating it, and reducing it a second time with dithionite, in the presence of 10 μ m cyanide. Cyanide suppresses the response of the interfering proteins to the second dithionite reduction. This modified method allows an accurate determination of the content of cytochrome b 559 using smaller numbers of cells than previously feasible.
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