Hapten-sandwich labeling. I. A general procedure for simultaneous labeling of multiple cell surface antigens for fluorescence and electron microscopy.
A hapten-sandwich procedure has been developed for specific labeling of cell surface antigens for fluorescence or electron microscopy. Haptens are azo-coupled to immunoglobulins specific for a cell surface antigen; the hapten-modified cell-bound antibodies can then be visualized by adding fluorescent antihapten antibody, or by adding antihapten antibody followed by hapten-modified markers for electron microscopy. Virus or high molecular weight protein markers are lightly cross-linked before conjugation with hapten to prevent their disruption. Such stable hapten-modified markers, and the accessibility of many different purified anti-azophenyl-hapten antibodies, make it feasible to distinguish more than one membrane antigen in a given labeling experiment. When mouse lymphoid cell populations are labeled with separate markers for Ig and for thymus-associated antigens, many cells exhibit the Ig marker exclusively or the thymic marker predominantly, and some cells are completely free of label.
D000818:Animals; D001152:Arsenicals; D001391:Azo Compounds; D001666:Binding Sites, Antibody; D002462:Cell Membrane / Q000276:immunology; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003979:Diazonium Compounds; D005260:Female; D005456:Fluorescent Dyes; D006027:Glycosides; D006041:Goats / Q000276:immunology; D006241:Haptens; D007074:Immunoglobulin G; D007136:Immunoglobulins; D007158:Immunologic Techniques; D007457:Iodine Radioisotopes; D007785:Lactose; D008214:Lymphocytes / Q000276:immunology; D008297:Male; D051379:Mice / Q000276:immunology; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008974:Mollusca / Q000276:immunology; D011817:Rabbits / Q000276:immunology; D014316:Tritium
D001152:Arsenicals; D001391:Azo Compounds; D003979:Diazonium Compounds; D005456:Fluorescent Dyes; D006027:Glycosides; D006241:Haptens; D007074:Immunoglobulin G; D007136:Immunoglobulins; D007457:Iodine Radioisotopes; D014316:Tritium; D007785:Lactose