Domain and basement membrane specificity of a monoclonal antibody against chicken type IV collagen.
A monoclonal antibody, IV-IA8, generated against chicken type IV collagen has been characterized and shown to bind specifically to a conformational-dependent site within a major, triple helical domain of the type IV molecule. Immunohistochemical localization of the antigenic determinant with IV-IA8 revealed that the basement membranes of a variety of chick tissues were stained but that the basement membrane of the corneal epithelium showed little, if any, staining. Thus, basement membranes may differ in their content of type IV collagen, or in the way in which it is assembled. The specificity of the antibody was determined by inhibition ELISA using purified collagen types I-V and three purified molecular domains of chick type IV collagen ([F1]2F2, F3, and 7S) as inhibitors. Only unfractionated type IV collagen and the (F1)2F2 domain bound the antibody. Antibody binding was destroyed by thermal denaturation of the collagen, the loss occurring at a temperature similar to that at which previous optical rotatory dispersion studies had shown melting of the triple helical structure of (F1)2F2. Such domain-specific monoclonal antibodies should prove to be useful probes in studies involving immunological dissection of the type IV collagen molecule, its assembly within basement membranes, and changes in its distribution during normal development and in disease.
D000818:Animals; D000911:Antibodies, Monoclonal; D001485:Basement Membrane / Q000032:analysis*; D001808:Blood Vessels / Q000032:analysis; D002642:Chick Embryo; D002645:Chickens; D003094:Collagen / Q000032:analysis* / Q000276:immunology; D004727:Endothelium / Q000032:analysis; D000939:Epitopes; D005123:Eye / Q000032:analysis; D005455:Fluorescent Antibody Technique; D007668:Kidney / Q000032:analysis; D009131:Muscle, Smooth, Vascular / Q000032:analysis; D009132:Muscles / Q000032:analysis; D009206:Myocardium / Q000032:analysis; D011487:Protein Conformation
D000911:Antibodies, Monoclonal; D000939:Epitopes; D003094:Collagen