CD1c (BDCA-1)+ Dendritic Cell Isolation Kit ( #130-091-878 )

CD1c (BDCA-1)⁺ Dendritic Cell Isolation Kit

Overview

The CD1c (BDCA-1)⁺ Dendritic Cell Isolation Kit includes FcR Blocking Reagent, CD1c (BDCA-1)-PE, Anti-PE MicroBeads for the enrichment of CD1c (BDCA-1)⁺ blood dendritic cells. CD20 MicroBeads are included for depletion of B cells prior to DC isolation.

Details

Background information
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: CD11c^highCD123^low CD1c (BDCA1)⁺ DCs and CD11c^dimCD123^– 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 and CD20 antibodies have been tested to cross-react with antigens of the cynomolgus monkey (Macaca fascicularis).

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.

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

$B cell–depleted flow-through fraction$

Counter-staining of B cell depleted flow-through fraction with CD14-APC

$Counter-staining of B cell depleted flow-through fraction with CD14-APC$

CD1c (BDCA-1)⁺ blood dendritic cells after enrichment

CD1c (BDCA-1)<sup>+</sup> blood dendritic cells after enrichment