Rapid replicate reverse haemolytic plaque assay directly in microwells used for tissue culture.
The reverse haemolytic plaque assay was adapted as a micromethod using microwells of flat-bottomed microtitre trays. To microwells containing the lymphocytes under test were added protein A-coupled ox erythrocytes, developing antisera directed against any class of immunoglobulin, and guinea pig complement absorbed with protein A-coupled erythrocytes. Plaques were scored by counting with a stereoscan microscope. The method is applicable both to spontaneous and mitogen-induced plaque formation and, as a further development of the technique to differentiated B cells in pokeweed mitogen-driven cultures, plaques were enumerated directly in the same wells of the microtitre tray as were employed for culture of the B cells. The method allows for several hundred plaque assays at one sitting.
D000921:Antibody-Producing Cells / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D006462:Hemolytic Plaque Technique; D006801:Humans; D008213:Lymphocyte Activation; D011043:Pokeweed Mitogens / Q000494:pharmacology
D011043:Pokeweed Mitogens