IMMUNOFLUORESCENT ANTIBODIES TO BOVINE KERATOHYALIN AND IMMUNOLOGIC CONFIRMATION OF HOMOLOGY

Abstract

Extraction of bovine hoof epidermis with 1.0 M potassium phosphate buffer (pH 7.0) and dialysis of the extract against distilled water results in the formation of macroaggregates. Fractionation of solubilized macroaggregates on polyacrylamide gels reveals a 13-member oligomeric series. Indirect immunofluorescence with antisera from rabbits immunized with either macroaggregates or gel slices with individual oligomers demonstrates specific fluorescence over keratohyaline granules. The reaction is tissue-specific, but antibodies cross-react with goat, sheep, wildebeest and rabbit keratohyaline granules, although not against human, rat or guinea pig keratohyalin. Immunofluorescent methods show that the macroaggregates are a component of keratohyalin and that the 13 oligomers are antigenically identical.