Differential inhibition of indirect immunofluorescence in rat liver sections incubated with antibodies against microsomal cytochromes P-450a, b, and c and epoxide hydrolase.
Rat liver sections were incubated with antibodies (100-1000 micrograms IgG/ml) against microsomal cytochromes P-450a, P-450b, and P-450c, and epoxide hydrolase. Inhibition of indirect immunofluorescence, which progressed with higher concentrations of primary antibody, corresponded with antigen-enriched tissue in frozen liver sections from male and female rats. It was found in liver sections from phenobarbital-treated rats incubated with anti-P-450b and anti-epoxide hydrolase and from 3-methylcholanthrene-treated rats incubated with anti-P-450c. No inhibition was found in sections from untreated rats or rats receiving treatments that did not induce the specific antigen. No inhibition was found in sections incubated with anti-P-450a. Inhibition of immunofluorescence was abolished in frozen sections subjected to dehydration-rehydration protocols known to extract antigens, and was prevented by certain solvents and detergent-wash. Inhibition of immunofluorescence provides a unique method for confirming the antigen-rich regions of the liver lobules specific for microsomal expoxide hydrolase and the cytochrome P-450s.
Moody
D E
DE
Department of Pathology, School of Medicine, University of California, San Francisco 94143.
Taylor
L A
LA
Smuckler
E A
EA
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000145:classification / Q000378:metabolism*; D004306:Dose-Response Relationship, Immunologic; D004851:Epoxide Hydrolases / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry / Q000379:methods; D008862:Microsomes, Liver / Q000201:enzymology* / Q000648:ultrastructure; D010634:Phenobarbital / Q000494:pharmacology; D051381:Rats
D003577:Cytochrome P-450 Enzyme System; D004851:Epoxide Hydrolases; D010634:Phenobarbital