| Overview |
| The Plasmacytoid Dendritic Cell Isolation Kit II allows the isolation of untouched human plasmacytoid dendritic cells (PDCs) from PBMCs. Having evolved from the first-generation kit (# 130-092-207), it now offers even better cell purity and yield in a faster procedure. |
| 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.
Detailed separation procedure
PDCs are isolated by depletion of non-PDCs. To this end, non-PDCs are magnetically labeled by using cocktails containing specific MicroBeads, biotin-conjugated antibodies, and Anti-Biotin MicroBeads. Magnetically labeled cells are retained in the column, while unlabeled PDCs with high purity are collected in the flow-through.
Downstream applications
PDCs isolated from human PBMCs can be used for phenotypic, functional, and molecular analyses, e.g., studies on expression of Toll-like receptors, chemokine receptors, or new antigens, and on DC activation, migration, cytokine production, and T cell polarization.
The Plasmacytoid Dendritic Cell Isolation Kit can also be used to enrich PDCs from tonsils, if the CD303 (BDCA-2) MicroBead Kit is not suitable. |
| Columns |
| LD, LS, or autoMACS Columns |
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| Figure 1 |
| Human PDCs were isolated from PBMCs by using the Plasmacytoid Dendritic Cell Isolation Kit II, an LD Column, and a MidiMACS™ Separator. Cells were stained with CD303 (BDCA-2)-APC and CD123-PE before or after separation and analyzed by flow cytometry using the MACSQuant Analyzer. |
| PBMCs before separation |
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| Enriched PDCs |
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