Immunohistochemical localization of collagens in the ear

The aim of this work was to prepare specific antibodies to various types of collagen in order to study on immunohistological localization of collagen under normal and pathologic conditions. Human type I and III collagens were extracted from human dura mater and placenta using limited pepsin digestion followed by differential salt precipitation and chromatographic purification. The purity of collagens was assessed by SDS polyacrylamide gel electrophoresis. Antibodies to type I and III were prepared in rabbits, and showed a high reaction with the corresponding human collagens, and these antibodies were tested to determine activity using ELISA. However, antibody to type I also showed some cross-reaction with type III, and antibody to type III, with type I. Using these antibodies, tissue distribution of collagen was examined in the middle ear by peroxidase anti-peroxidase method. In granulation tissue, it was revealed that types I and III co-existed.