Separation and detection of nitroblue tetrazolium-reducing enzymes from human polymorphonuclear leukocytes

A technique for detecting the nitroblue tetrazolium-reducing activity of human polymorphonuclear leukocyte enzymes separated on polyacrylamide gels is reported. Slices of gel are incubated in buffer containing nitroblue tetrazolium and either NADPH or NADH. Enzymes capable of reducing nitroblue tetrazolium form a dark band due to the formation of the insoluble formazan. By this technique the soluble fraction of human polymorphonuclear leukocyte sonicates was shown to contain several enzymes capable of reducing nitroblue tetrazolium in the presence of NADPH and NADH.