Separation of lymphocyte subpopulations using biotin-avidin erythrocyte rosettes.
The major method for isolating murine lymphocyte subpopulations involves negative selection using antibody plus complement-mediated cytolysis. We have developed an efficient rosette method for enriching murine B and T cells using biotin-conjugated antibodies and avidin-coated sheep erythrocytes. Rosetted and non-rosetted subpopulations are separated rapidly on Percoll cushions. In systems employing rabbit anti-mouse immunoglobulin or monoclonal rat anti-mouse Thy-1.2 conjugated to biotin, positively-selected cells are greater than 90% pure while negatively-depleted populations possess less than 2% contamination with unwanted cells. Recoveries from starting spleen cell populations range between 50 and 75%. This method provides an easily performed alternative for obtaining positively and negatively selected cell populations and can be used with any biotin-conjugated antibody protein.
D000818:Animals; D001360:Avidin; D001402:B-Lymphocytes / Q000276:immunology; D001710:Biotin; D002469:Cell Separation / Q000379:methods*; D004912:Erythrocytes; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D012397:Rosette Formation; D012756:Sheep; D013154:Spleen / Q000166:cytology; D013601:T-Lymphocytes / Q000276:immunology
D001360:Avidin; D001710:Biotin