| Overview |
Please notice: This product has been replaced by the Diamond Plasmacytoid Dendritic Cell Isolation Kit II, human (# 130-097-240), which has even better performance.
The Diamond Plasmacytoid Dendritic Cell Isolation Kit combines the isolation of untouched plasmacytoid dendritic cells (PDCs) by depletion of non-PDCs with the subsequent positive selection using MicroBeads against the 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 almost pure PDCs. |
| Details |
Background information
The isolated PDCs are CD303 (BDCA-2)+, CD304 (BDCA-4/Neuropilin-1)+, CD123+, CD4+, CD45RA+, CD141 (BDCA-3)dim, CD1c (BDCA-1)–, and CD2–. The cells lack expression of lineage markers (CD3, CD14, CD16, CD19, CD20, CD56) and express neither myeloid markers, such as CD13 and CD33, nor Fc receptors, such as CD16, CD64, or FcεRI.
Binding of the antibody to CD304 (BDCA-4/Neuropilin-1) does not have a significant 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
Detailed separation procedure
For the first step, all non-PDC cells are magnetically labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads for depletion over a MACS Column. In the subsequent positive selection step, the enriched PDCs are directly magnetically labeled with CD304 (BDCA-4/Neuropilin-1) MicroBeads.
Downstream applications
The Diamond Plasmacytoid Dendritic Cell Isolation Kit was developed for the isolation of maximum-purity PDCs from PBMCs. Therefore, it is particularly useful if almost pure PDCs are needed as starting material, e.g. for 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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