A new method to detect directly in culture cell surface membrane immunoglobulins.
An ELISA assay is described for the measurement of the smIgG. The method is based on the detection of cell-smIgG directly on the same microplate used for the culture. The cells, preincubated at 37 degrees C for one hour, were cultured in the presence of S-ConA and serum-free medium for two days. Using this strategy, the background noise due to non specific adsorption of IgG to plastic wells and cytophilic antibodies was eliminated. The cells in the presence of S-ConA and serum-free medium adhered to the plastic wells, and the cell-smIgG were detected using an anti-human IgG covalently linked to alkaline phosphatase or its F(ab')2 fragment. The possibility of measuring the modulation of the expression of the cell-smIgG without any additional manipulation is stressed.
D000328:Adult; D001402:B-Lymphocytes / Q000276:immunology*; D002462:Cell Membrane / Q000276:immunology; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D008297:Male; D011947:Receptors, Antigen, B-Cell / Q000032:analysis*; D013601:T-Lymphocytes / Q000276:immunology*
D007074:Immunoglobulin G; D011947:Receptors, Antigen, B-Cell