An ELISA for screening hybridoma cultures for monoclonal antibodies against a detergent solubilized integral membrane protein.
A method is described for the binding of a detergent solubilized integral membrane protein to polystyrene immunoassay plates. Addition of Bouin's fluid, a histochemical fixative, to wells of plates containing the detergent solubilized antigen, followed by low speed centrifugation, is sufficient to promote binding of antigen in the presence of Triton X-100 concentrations as high as 1.75%. The binding of antigen is rapid and the entire binding procedure, including removal of fixative and washing of the plates, can be accomplished in less than 15 min. Immunological specificity of the bound antigen is retained. This method has been used to effectively screen hybridoma cultures for specific antibodies.
D000085:Acetates / Q000494:pharmacology; D019342:Acetic Acid; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D001665:Binding Sites / Q000187:drug effects; D003902:Detergents; D003576:Electron Transport Complex IV / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005404:Fixatives; D005557:Formaldehyde / Q000494:pharmacology; D006825:Hybridomas / Q000276:immunology*; D007106:Immune Sera / Q000494:pharmacology; D007124:Immunoenzyme Techniques; D008099:Liver / Q000201:enzymology; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D017830:Octoxynol; D010851:Picrates / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology*; D051381:Rats
D000085:Acetates; D000911:Antibodies, Monoclonal; D003902:Detergents; D005404:Fixatives; D007106:Immune Sera; D008565:Membrane Proteins; D010851:Picrates; D005557:Formaldehyde; C026239:Bouin's solution; D011092:Polyethylene Glycols; D017830:Octoxynol; D003576:Electron Transport Complex IV; D019342:Acetic Acid