Monoclonal antibodies against the mercaptoethanol-sensitive structure of a cell-cell adhesion protein of Polysphondylium pallidum.
Monoclonal antibodies were prepared against a putative cell-cell adhesion glycoprotein, with an apparent molecular mass of 64,000 (gp64), of the cellular slime mold, Polysphondylium pallidum. Five monoclonal antibodies obtained by means of an enzyme-linked immunoadsorbent assay did not bind to the antigens which were subjected to gel electrophoresis and blotting method in the presence of a reducing agent, but they did bind specifically to the antigens prepared in unreducing conditions of samples and then processed by the same blotting method. To solubilize gp64 in a sodium dodecyl sulfate (SDS)-sample buffer without mercaptoethanol (heated) or SDS-sample buffer with 2-mercaptoethanol (nonheated) was critical for the antibody binding onto gp64 on a membrane. Hence the antibodies seem to bind to a surface portion(s) of the localized protein structure folded up by disulfide cross-linkages. One of the antibodies obtained blocked cell-cell adhesion by about 20%.
Manabe
R
R
Department of Botany, Faculty of Science, Hokkaido University.
Manabe
N
N
Ochiai
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000946:Antigens, Fungal / Q000276:immunology; D001667:Binding, Competitive; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects*; D015815:Cell Adhesion Molecules / Q000276:immunology*; D002462:Cell Membrane / Q000648:ultrastructure; D006023:Glycoproteins / Q000276:immunology*; D006358:Hot Temperature; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009235:Myxomycetes / Q000187:drug effects / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D008623:Mercaptoethanol