Differential inhibition of indirect immunofluorescence in rat liver sections incubated with antibodies against microsomal cytochromes P- 450a, b, and c and epoxide hydrolase

• Published in: The journal of histochemistry and cytochemistry Vol. 36; no. 3; pp. 291 - 295
• Main Authors: Moody, DE, Taylor, LA, Smuckler, EA
• Format: Journal Article
• Language: English
• Published: Los Angeles, CA Histochemical Soc 01-03-1988; SAGE Publications; Histochemical Society
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Cytochrome P-450 Enzyme System - classification; Cytochrome P-450 Enzyme System - metabolism; Dose-Response Relationship, Immunologic; Enzymes and enzyme inhibitors; Epoxide Hydrolases - metabolism; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Immunohistochemistry - methods; Microsomes, Liver - enzymology; Microsomes, Liver - ultrastructure; Oxidoreductases; Phenobarbital - pharmacology; Rats; Immunohistochemistry; Hemoprotein; Rat; Antibody; Cytochrome P450; Liver; Rodentia; Microsome; Epoxide hydrolase; Vertebrata; Mammalia; Flavoprotein-linked monooxygenase; Inhibition; Immunofluorescence
• ISSN: 0022-1554; 1551-5044
• DOI: 10.1177/36.3.3278056

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.