LUMINESCENT VISUALIZATION OF LOW AMOUNTS OF CYTOCHROME-P450 AND HEMOPROTEINS BY LUMINOL IN ACRYLAMIDE GELS

In this paper we apply luminol (5-amino-2,3-dihydro-1,4-phthalazinedio ne), a light-emitting substrate, in conjunction with H2O2 to the lumin escence labeling of hemoproteins. We describe in detail a photodetecti on device which permits an efficient recording of the light emitted by heme-containing proteins resolved in acrylamide gels. The sensitivity of this procedure, when compared to the classical 3,3'-5,5'-tetrameth ylbenzidine staining method, results in a 5- to 20-fold enhancement wi th standard hemoproteins (lactoperoxidase, catalase, cytochrome P450 2 B4, horseradish peroxidase, hemoglobin, myoglobin, and cytochrome b5). The potential applications of this technique are illustrated by the d etection of cytochrome P450 in microsomes from plant as well as from a nimal extrahepatic tissues which possess low amounts of this cytochrom e. (C) 1995 Academic Press, Inc.