A rapid and efficient method for purification of rat B cells for complement-dependent cytotoxicity testing.
The development of an effective technique for the enrichment of rat B cells is described. This protocol is an adaptation of the 'panning' technique employing anti-Ig-coated plastic dishes. Rat B cells are bound directly to anti-rat Ig-coated wells in Terasaki test plates where they are typed by complement-dependent cytotoxicity. This technique enriches class II (Ia-like) antigen-positive B cells, facilitating detection with antisera specific for rat class II alloantigens. The utility of this technique in the cell surface phenotyping of minor lymphocyte subpopulations is discussed.
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D003165:Complement System Proteins / Q000378:metabolism*; D003601:Cytotoxicity Tests, Immunologic; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007106:Immune Sera / Q000494:pharmacology; D051379:Mice; D008805:Mice, Inbred A; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D051381:Rats; D011914:Rats, Inbred BN; D011915:Rats, Inbred BUF; D011916:Rats, Inbred F344; D011917:Rats, Inbred Lew; D011920:Rats, Inbred WF
D000949:Histocompatibility Antigens Class II; D007106:Immune Sera; D003165:Complement System Proteins