BD IMag™ Mouse Dendritic Cell Enrichment Set - DM

Enrichment of DC from mouse spleen. BALB/c splenocytes were labeled with the BD IMag™ Mouse Dendritic Cell Enrichment Set - DM 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 Anti-Mouse CD11c HL3 (Cat. No. 557400/553801) to detect DC and a mixture of PE Anti-Mouse CD2 RM2-5 (Cat. No. 553112), Anti-Mouse CD3e 145-2C11 (Cat. No. 553063/553064), Anti-Mouse CD45R/B220 RA3-6B2 (Cat. No. 553089/553090), Anti-Mouse CD49b HMa2 (Cat. No. 558759), Anti-Mouse Ly6-G and Ly-6C RB6-8C5 (Cat. No. 553128), and Anti-Mouse TER119/Erythroid Cells TER-119 (Cat. No. 553673) monoclonal antibodies to detect non-DC 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 DC is indicated in the lower-right corner of each panel. Left panel shows unseparated splenocytes.  Middle panel shows the twice-enriched fraction (after two 6-minute magnetic separations plus an additional 6-minute separation of those enriched cells) from splenocytes that were not treated with BD Fc Block™ antibody (Cat. No. 553142/553141).  Right panel shows the twice-enriched fraction from splenocytes that were treated with BD Fc Block™ antibody.  The addition of the BD Fc Block™ antibody increases the DC recovery by about 15-25%, while it reduces the purity of the enriched fraction by 7-15%.