| Description |
The Diamond Plasmacytoid Dendritic Cell Isolation Kit combines isolation of untouched plasmacytoid dendritic cells (PDCs) by depletion of non-PDCs and their subsequent positive selection using MicroBeads against PDC-specific antigen CD304 (BDCA-4/Neuropilin-1). By this two-step magnetic separation procedure, the purity of the isolated PDC population, which is already high after depletion of non-PDCs, is further increased, resulting in an almost pure PDC fraction. For depletion, non-PDCs (i.e. T cells, B cells, NK cells, myeloid dendritic cells, monocytes, granulocytes, and erythroid cells) are magnetically labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. For subsequent positive selection, the enriched PDCs are directly magnetically labeled with CD304 (BDCA-4/Neuropilin-1) MicroBeads. Binding of the antibody to CD304 (BDCA-4/Neuropilin-1) does not have a substantial effect on IFN type I production in PDCs, as it was shown for binding of antibodies to the other PDC-specific antigen CD303 (BDCA‑2).1, 2
The Diamond Plasmacytoid Dendritic Cell Isolation Kit was developed for the isolation of highly pure PDCs from PBMCs. It includes the Non-PDC Biotin-Antibody Cocktail, Anti-Biotin MicroBeads, and CD304 (BDCA-4/Neuropilin-1) Diamond MicroBeads.
For evaluation of MACS® Separations, staining with CD303 (BDCA-2) antibodies is recommended. |
| Applications |
| The Diamond Plasmacytoid Dendritic Cell Isolation Kit is particularly useful if almost pure PDCs are needed as starting material, for example gene expression profiling or protein purification. |
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| Figure 1 |
Isolation of PDCs from PBMCs using the Diamond Plasmacytoid Dendritic Cell Isolation Kit, an LD Column and a MidiMACS™ Separator, as well as a MiniMACS™ Separator and two MS Columns.
Cells were stained with CD303 (BDCA-2)-APC and CD123-PE. Like CD304 (BDCA-4/Neuropilin-1), the CD303 (BDCA-2) antigen is specifically expressed on plasmacytoid dendritic cells in blood and allows their direct identification. |
| A: PBMCs before separation |
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| B: Untouched PDCs after depletion of non-PDCs |
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| C: PDCs after positive selection with CD304 (BDCA-4/Neuropilin-1) MicroBeads |
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| Figure 2 |
| Isolated PDCs were cultured for 48 hours in medium with IL-3 and CpG B, and subsequently stained for expression of CD63 (green) and HLA-DR (red). |
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