Immunogold localization of peroxisomal enzymes in Epon-embedded liver tissue. Enhancement of sensitivity by etching with ethanolic sodium hydroxide.
Epon-embedded biological materials exhibit well preserved ultrastructure but generally weak antigenicity. Brief etching of ultrathin Epon sections with resin solvent, ethanolic sodium hydroxide, brought about a nearly 2-fold increase in the immunogold labeling density of rat and human hepatic peroxisomes after incubation with antisera against catalase and 3 enzymes of lipid beta-oxidation: acyl-CoA oxidase, bifunctional protein (enoyl-CoA hydratase-3-hydroxyacyl-CoA dehydrogenase) and 3-ketoacyl-CoA thiolase. The etching was superior to pretreatment with an oxidant, sodium metaperiodate. Despite some deterioration of the cellular ultrastructure, the obtained enhancement of the sensitivity of the protein A-gold method may be helpful in cases when the antigenicity to be detected is strongly inhibited by epoxy resin embedding.
Litwin
J A
JA
Institute of Anatomy, University of Heidelberg, Federal Republic of Germany.
Beier
K
K
D000102:Acetyl-CoA C-Acyltransferase / Q000032:analysis; D042962:Acyl-CoA Oxidase; D000818:Animals; D002374:Catalase / Q000032:analysis; D004746:Enoyl-CoA Hydratase / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000201:enzymology* / Q000648:ultrastructure; D008830:Microbodies / Q000201:enzymology*; D008854:Microscopy, Electron; D010088:Oxidoreductases / Q000032:analysis; D010504:Periodic Acid; D051381:Rats; D012972:Sodium Hydroxide; D013205:Staphylococcal Protein A
D013205:Staphylococcal Protein A; D010504:Periodic Acid; D012972:Sodium Hydroxide; C009288:metaperiodate; D010088:Oxidoreductases; D002374:Catalase; D042962:Acyl-CoA Oxidase; D000102:Acetyl-CoA C-Acyltransferase; D004746:Enoyl-CoA Hydratase