A simple method for detecting anti-tubular membrane antibodies in rat lymphoid cell cultures.
The application of enzyme-linked immunosorbent assay (ELISA) to short cell cultures has proved to be useful in detecting immunoglobulins secreted to supernatants. This paper describes a modified ELISA to detect and quantify the production of specific anti-tubular basal membrane IgG released in vitro by lymphoid cells from Brown Norway rats with tubulointerstitial nephritis. This method uses 4-methyl-umbelliferyl-phosphate as substrate and allows to detect approximately concentrations 100 times less than the detectable concentration of visibly colored substrates. This method eliminates the need for mitogens, is precise and reproducible and the use of 4-methyl-umbelliferyl-phosphate allows a substantial increase in sensitivity.
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000954:Antigens, Surface / Q000276:immunology*; D001485:Basement Membrane / Q000276:immunology*; D002417:Cattle; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains
D000906:Antibodies; D000954:Antigens, Surface