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Enumeration of antigen-specific IgE responses at the single-cell level by an ELISA plaque assay
1Department of Pathology, Case Western Reserve University, Cleveland, OH 44106.
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.
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.