The Pan-DC Enrichment Kit is a fast pre-enrichment kit for untouched isolation of plasmacytoid and myeloid dendritic cells from PBMCs.

Data and images for Pan-DC Enrichment Kit, human

Figures

Figure 1

Pre-enrichment of untouched Pan-DCs from human PBMCs using the Pan-DC Enrichment Kit, human, a MidiMACS Separator and an LS Column. The cells were fluorescently stained with CD303 (BDCA-2)-FITC, CD141 (BDCA-3)-APC, CD1c (BDCA-1)-PE, and CD20-PerCP and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and B cells were excluded from the analysis based on scatter signals, propidium iodide fluorescence, and CD20 expression.
Unseparated fraction
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Figure 1

Pre-enrichment of untouched Pan-DCs from human PBMCs using the Pan-DC Enrichment Kit, human, a MidiMACS Separator and an LS Column. The cells were fluorescently stained with CD303 (BDCA-2)-FITC, CD141 (BDCA-3)-APC, CD1c (BDCA-1)-PE, and CD20-PerCP and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and B cells were excluded from the analysis based on scatter signals, propidium iodide fluorescence, and CD20 expression.
View details

Figure 1

Pre-enrichment of untouched Pan-DCs from human PBMCs using the Pan-DC Enrichment Kit, human, a MidiMACS Separator and an LS Column. The cells were fluorescently stained with CD303 (BDCA-2)-FITC, CD141 (BDCA-3)-APC, CD1c (BDCA-1)-PE, and CD20-PerCP and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and B cells were excluded from the analysis based on scatter signals, propidium iodide fluorescence, and CD20 expression.
Enriched Pan-DCs
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Figure 1

Pre-enrichment of untouched Pan-DCs from human PBMCs using the Pan-DC Enrichment Kit, human, a MidiMACS Separator and an LS Column. The cells were fluorescently stained with CD303 (BDCA-2)-FITC, CD141 (BDCA-3)-APC, CD1c (BDCA-1)-PE, and CD20-PerCP and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and B cells were excluded from the analysis based on scatter signals, propidium iodide fluorescence, and CD20 expression.
View details

Figure 1

Pre-enrichment of untouched Pan-DCs from human PBMCs using the Pan-DC Enrichment Kit, human, a MidiMACS Separator and an LS Column. The cells were fluorescently stained with CD303 (BDCA-2)-FITC, CD141 (BDCA-3)-APC, CD1c (BDCA-1)-PE, and CD20-PerCP and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and B cells were excluded from the analysis based on scatter signals, propidium iodide fluorescence, and CD20 expression.
Non-DCs
View details

Figure 1

Pre-enrichment of untouched Pan-DCs from human PBMCs using the Pan-DC Enrichment Kit, human, a MidiMACS Separator and an LS Column. The cells were fluorescently stained with CD303 (BDCA-2)-FITC, CD141 (BDCA-3)-APC, CD1c (BDCA-1)-PE, and CD20-PerCP and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and B cells were excluded from the analysis based on scatter signals, propidium iodide fluorescence, and CD20 expression.
View details

Figure 1

Pre-enrichment of untouched Pan-DCs from human PBMCs using the Pan-DC Enrichment Kit, human, a MidiMACS Separator and an LS Column. The cells were fluorescently stained with CD303 (BDCA-2)-FITC, CD141 (BDCA-3)-APC, CD1c (BDCA-1)-PE, and CD20-PerCP and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and B cells were excluded from the analysis based on scatter signals, propidium iodide fluorescence, and CD20 expression.

Specifications for Pan-DC Enrichment Kit, human

Overview

The Pan-DC Enrichment Kit is a fast pre-enrichment kit for untouched isolation of plasmacytoid and myeloid dendritic cells from PBMCs.

Detailed product information

Detailed separation procedure

Using the Pan-DC Enrichment Kit, dendritic cells (MDC1s, MDC2s, and PDCs) are isolated very fast in a single step by depletion of non-target cells (negative selection). Non-target cells are labeled with a cocktail of biotin-conjugated monoclonal antibodies and Anti-Biotin MicroBeads. Additionally, they are directly magnetically labeled with a cocktail of MicroBead-conjugated antibodies against antigens that are not expressed by dendritic cells. The magnetically labeled non-target cells are depleted by retaining them within a MACS Column in the magnetic field of a MACS Separator while the unlabeled DCs run through.

Downstream applications

Dendritic cells pre-enriched with the Pan-DC Enrichment Kit can be used for:
  • Analysis of antigen uptake, processing, and presentation
  • Stimulation of primary T cell responses by antigen-pulsed dendritic cells
  • Analysis of gene expression in blood dendritic cells
  • Elucidation of specific signal transduction pathways

Columns

LS, XS, or autoMACS
®
Columns.

References for Pan-DC Enrichment Kit, human

Publications

  1. O’Doherty, U. et al. (1994) Human blood contains two subsets of dendritic cells, one immunologically mature and the other immature. Immunology 82: 487-493
  2. Dzionek, A. et al. (2000) BDCA-2, BDCA-3, BDCA-4: Three markers for distinct subsets of dendritic cells in human peripheral blood. J. Immunol. 165: 6037-6046
  3. Penna, G. et al. (2001) Cutting Edge: Selective usage of chemokine receptors by plasmacytoid dendritic cells.. J. Immunol. 167: 1862-1866
  4. Penna, G. et al. (2002) Cutting edge: differential chemokine production by myeloid and plasmacytoid dendritic cells. J. Immunol. 169: 6673-6676
  5. Krug, A. et al. (2001) Toll-like receptor reveals CpG DNA as a unique microbial vstimulus for plasmacytoid dendritic cells which synergizes with CD40 ligand to induce high amounts of IL-12. Eur. J. Immunol. 31: 3026-3037

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