Monoclonal antibodies that distinguish avian type I and type III collagens: isolation, characterization and immunolocalization in various tissues.
Monoclonal antibodies were prepared that were specific for chicken type I and type III collagens. The specificity of these antibodies was determined by ELISA, inhibition ELISA, and immunoblot assays. The results showed that the monoclonal antibodies were specific for their respective antigens without significant cross reactivity to other types of collagen. An analysis of the location of the epitopes by rotary shadowing that a monoclonal antibody for type I collagen (called DD4) recognized type I procollagen close to the large globular domain at the carboxyl terminus of the molecule. A monoclonal antibody for type III collagen (called 3B2) recognized both the intact type III molecule and also the TCA fragment of type III collagen after mammalian collagenase digestion. The epitope was located approximately one-fifth of the distance from the amino-terminus of the intact molecule. The monoclonal antibodies were used for immunolocalization of type I and type III collagens in cryosections of heart, aorta, kidney, liver, thymus, skin, gizzard and myotendinous junction. In heart, aorta, kidney, liver, thymus and skin, type I and III collagens were colocalized in the connective tissue of each organ. In contrast, gizzard and myotendinous junction showed distinctly different staining patterns for the distribution of type I and type III collagen. The two monoclonal antibodies reported here are potentially useful reagents to study fibril formation involving type I and type III collagens.
Swasdison
S
S
Department of Cell Biology, University of Alabama, Birmingham 35294.
Mayne
P M
PM
Wright
D W
DW
Accavitti
M A
MA
Fitch
J M
JM
Linsenmayer
T F
TF
Mayne
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001011:Aorta / Q000737:chemistry; D002645:Chickens / Q000276:immunology; D003094:Collagen / Q000032:analysis / Q000145:classification / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D005895:Gizzard, Avian / Q000737:chemistry; D012867:Skin / Q000737:chemistry; D014781:Viscera / Q000737:chemistry
D000911:Antibodies, Monoclonal; D003094:Collagen