BD IMag™ Mouse CD4 T Lymphocyte Enrichment Set - DM

Enrichment of CD4 T lymphocytes from mouse spleen. BALB/c splenocytes were labeled with the BD IMag Mouse CD4 T Lymphocyte Enrichment Set - DM (Cat. No. 558131) and separated on the BD IMag™ Cell Separation Magnet (Cat. No. 552311) according to the accompanying protocol. To demonstrate the efficiency of the enrichment, cells were stained with FITC Rat Anti-Mouse CD4 (Cat. No. 557307/553729) to detect CD4 T lymphocytes and a mixture of PE Rat Anti-Mouse CD8a (Cat. No. 553032/553033), CD11b (Cat. No. 557397/553311), CD45R/B220 (Cat. No. 553089/553090), CD49b (Cat. No. 558759), and TER-119/Erythroid Cells (Cat. No. 553673) monoclonal antibodies to detect CD4-negative leukocytes and erythrocytes. Dead cells were excluded by staining with Propidium Iodide Staining Solution (Cat. No. 556463). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system. Please refer to the Enrichment Flow Chart to identify the cell populations represented in this figure. The percentage of CD4 T cells is indicated in the lower-right corner of each panel. Far left panel shows unseparated splenocytes. Middle left panel shows the combined enriched fraction after three 6-minute magnetic separations. Middle right panel shows the twice-enriched fraction after an additional 6-minute separation of the cells shown in middle left panel. This additional incubation can result in up to 5% increased purity with up to a 5% decrease in recovery. Far right panel shows the positive fraction.