Enumeration of antigen-specific IgE responses at the single-cell level by an ELISA plaque assay.
In vitro elicitation and enumeration of antigen-specific IgE-secreting cells in antigen-primed murine lymphocyte cultures can be reproducibly and accurately quantitated. Rat anti-mouse IgE monoclonal antibodies (Mab anti-epsilon) were developed. Antigen-specific IgE-secreting cells can be detected on antigen or MAb anti-epsilon coated nitrocellulose discs pasted to 24-well culture plates. This IgE ELISA-plaque assay (EP epsilon) is more reliable than antigen capture or total IgE ELISAs in monitoring de novo induction of antigen-specific IgE memory responses in cultures of antigen-primed lymphocytes elicited with antigens.
Chen
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Department of Pathology, Case Western Reserve University, Cleveland, OH 44106.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000921:Antibody-Producing Cells / Q000276:immunology*; D001710:Biotin; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D005260:Female; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D007073:Immunoglobulin E / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D007132:Immunoglobulin Isotypes / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011863:Radioimmunoassay; D051381:Rats
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007132:Immunoglobulin Isotypes; D007073:Immunoglobulin E; D001710:Biotin