Generation of monoclonal antibodies specific for developmentally regulated antigens of the chicken retina.
In order to raise monoclonal antibodies specific for neural antigens that might be involved in regulatory events during histogenesis of the chick visual system, 5 different immunogen fractions were employed in conjunction with 4 immunization regimes. By screening 17,000 hybridoma supernatants from 58 fusion experiments, empirical guidelines were formulated for generation of antibodies with distinct specificities. A number of positive and negative results would not have been predicted from theoretical considerations and emphasize the relevance of empirical data. To substantiate our observations we present 10 monoclonal antibodies that bind to distinct cell types or layers of the retina. Single cell as well as explant cultures in conjunction with Western blot analysis were employed to determine subcellular localization and cell type specificity of selected antibodies. Six antibodies bind specifically to the inner limiting membrane or the optic fiber layer or the inner plexiform layer or the outer plexiform layer. Four recognize selectively ganglion cells or Müller glial cells or presumably subpopulations of amacrine cells. These antibodies promise to be helpful tools to evaluate mechanisms of neural differentiation.
Max-Planck-Institut für Entwicklungsbiologie, Tübingen, F.R.G.
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Schlosshauer
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D002459:Cell Fusion; D002478:Cells, Cultured; D002645:Chickens; D002846:Chromatography, Affinity; D003429:Cross Reactions; D003520:Cyclophosphamide / Q000494:pharmacology; D003599:Cytoskeleton / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006054:Goldfish / Q000276:immunology; D037102:Lectins; D051379:Mice; D009457:Neuroglia / Q000276:immunology; D009474:Neurons / Q000276:immunology; D051381:Rats; D012160:Retina / Q000254:growth & development* / Q000276:immunology*; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000941:Antigens; D037102:Lectins; D003520:Cyclophosphamide