Purification and Characterization of Canine Collagens I and II

Abstract

The purpose of this study was to demonstrate that commercially available human anti-collagen antibodies are suitable for use with canine tissues. This paper describes a reproducible procedure for isolating and purifying canine skin and cartilage collagens in quantities sufficient to characterize. The skin and cartilage were readily dissolved in dilute acetic acid with pepsin, and the collagens were purified by salt precipitation. Purified collagens were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by immunodetection on electroblot. The tissue distribution of the canine collagens was examined by immunohistochemistry. Two polypeptide chains were identified by SDS-PAGE in the canine skin sample and a human collage Type I control. One polypeptide chain was identified in the canine articular cartilage sample and a human collagen Type II control. Immunoblotting revealed the individual components for the collagen antigens. Anti-human Type II collagen immunoreacted specifically with cells of a chondrocytic phenotype. Antihuman Type I collagen appeared to be less specific. Because the antibodies exhibited cross-species reactivity, homology between canine Type I and II collagens and human Type I and II collagens is suggested. Commercial antibodies for human collagens Type I and II are suitable for study of these canine collagens. (The J Histotechnol 24:51, 2001)Submitted: April 5, 2000; Accepted with revisions: January 5, 2001