Abstract
By means of a combined histochemical-immunological technique, at least a part of the intracellular hepatic cytochrome P448 has been localized within nuclei isolated from 3-methylcholanthrene pretreated rats. In this technique, sections of nuclei are incubated initially with rabbit anti-cytochrome P448, then with goat anti-rabbit immunoglobulin to which β-galactosidase had been chemically coupled. The preparation is then histochemically assayed for β-galactosidase using the histochemical substrate 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside in the presence of an oxidizing agent, nitroblue tetrazolium. A blue pigment is generated at the site of localization of β-galactosidase, and accordingly, of cytochrome P448. When conjugate was employed in which the immunoglobulin was monospecific antibody to cytochrome P450 obtained from the livers of phenobarbital-treated rats, no demonstrable activity was observed within the nuclei obtained from 3-methylcholanthrene pretreated rats. This control adds considerable weight to our argument for the intranuclear localization of at least a portion of the cytochrome P448.
- Copyright © 1979 by Academic Press, Inc.
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