Immunocytochemical staining of rat tissues with antibodies to denatured type I collagen: a technique for localizing areas of collagen degradation.
Sheep were immunized with purified native collagen I and the antisera were fractionated by affinity chromatography on columns containing either native or heat-denatured forms of the antigen. Crossed immunoadsorption ensured complete removal of antibodies to terminal, non-helical regions resulting in antibodies that were specific for either native or central, non-helical determinants, as judged by ELISA and immunoblotting. Tissue immunostaining showed some areas, such as duodenal villus and blood vessels, where staining with antibodies against native and denatured collagen gave similar patterns, but other areas, such as pancreas, where immunolocalization of native collagen I was not accompanied by similar detection of the denatured form. Immunodetection on tissue sections of added human fibronectin showed similar staining patterns to those for denatured collagen. Areas of tissue showing the presence of denatured collagen were interpreted as an indication of collagen degradation, and the technique may therefore provide a means of assessing the pattern of collagen turnover within individual tissues.
D000818:Animals; D000906:Antibodies; D002846:Chromatography, Affinity; D003094:Collagen / Q000032:analysis*; D004386:Duodenum / Q000166:cytology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005353:Fibronectins / Q000276:immunology; D006801:Humans; D008970:Molecular Weight; D010179:Pancreas / Q000166:cytology*; D011489:Protein Denaturation; D051381:Rats; D012867:Skin / Q000032:analysis
D000906:Antibodies; D005353:Fibronectins; D003094:Collagen