CD1c (BDCA-1)+ Dendritic Cell Isolation Kit
Description Human as well as rhesus monkey dendritic cells represent a heterogenous population composed of several subsets. Myeloid and plasmacytoid DCs, differentiated by their expression of CD11c and CD123, respectively, were further characterized by means of new markers against human blood dendritic cell antigens (BDCA). Two human myeloid dendritic cell populations have been described: CD11chigh CD123low CD1c (BDCA1)+ DCs and CD11cdim CD123– CD141 (BDCA-3)+ DCs. Plasmacytoid CD11c– CD123+ dendritic cells are characterized by the expression of CD303 (BDCA-2) or CD304 (BDCA-4).1 In rhesus monkey, CD1c is expressed on myeloid CD11c+ CD123– dendritic cells2 as well as on a subset of B cells. The CD1c (BDCA-1)+ Dendritic Cell Isolation Kit includes FcR Blocking Reagent, CD1c (BDCA-1)-PE, Anti-PE MicroBeads, and CD20 MicroBeads for depletion of B cells prior to the enrichment of CD1c (BDCA-1)+ blood dendritic cells. Applications The CD1c (BDCA-1)+ Dendritic Cell Isolation Kit can be used for the separation of CD1c+ dendritic cells from rhesus monkey (Macaca mulatta) PBMCs. The CD1c and CD20 antibodies have been tested to cross-react with antigens of the cynomolgus monkey (Macaca fascicularis). Columns For the first magnetic separation (depletion): LD or autoMACS™ Columns. For the following magnetic separation (positive selection): MS, LS, or autoMACS Columns.		Figure 1 Isolation of CD1c (BDCA-1)+ dendritic cells from rhesus monkey PBMCs using the CD1c (BDCA-1)+ Dendritic Cell Isolation Kit. Cells were incubated with CD1c (BDCA-1)-PE and subsequently magnetically labeled with CD20 MicroBeads. B cells were depleted using an LD Column. The B cell depleted fraction was magnetically labeled with Anti-PE MicroBeads and separated over an MS Column. Cell samples were stained with CD20-FITC and CD14-APC. Cell debris and dead cells were excluded from the analysis based on scatter signals and PI fluorescence. PBMCs before separation B cell–depleted flow-through fraction Counter-staining of B cell depleted flow-through fraction with CD14-APC CD1c (BDCA-1)+ blood dendritic cells after enrichment