The Diamond Plasmacytoid Dendritic Cell Isolation Kit II yields PDC populations with excellent purities. The kit involves a two-step procedure, consisting of depletion of non-PDCs and positive selection using MicroBeads against the PDC-specific antigen CD304 (BDCA-4/Neuropilin-1). This kit has been developed based on the first-generation kit (# 130-092-402) for even better performance and a shorter protocol.

Data and images for Diamond Plasmacytoid Dendritic Cell Isolation Kit II, human

Figures

Figure 1

Isolation of PDCs from PBMCs using the Diamond Plasmacytoid Dendritic Cell Isolation Kit II, 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 PDCs in blood and allows their direct identification.
PBMCs before separation
Enriched PDCs after depletion of non-PDCs
View details

Figure 1

Isolation of PDCs from PBMCs using the Diamond Plasmacytoid Dendritic Cell Isolation Kit II, 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 PDCs in blood and allows their direct identification.
View details

Figure 1

Isolation of PDCs from PBMCs using the Diamond Plasmacytoid Dendritic Cell Isolation Kit II, 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 PDCs in blood and allows their direct identification.
Pure PDCs after positive selection with CD304 (BDCA-4/Neuropilin-1) Diamond MicroBeads
View details

Figure 1

Isolation of PDCs from PBMCs using the Diamond Plasmacytoid Dendritic Cell Isolation Kit II, 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 PDCs in blood and allows their direct identification.

Specifications for Diamond Plasmacytoid Dendritic Cell Isolation Kit II, human

Overview

The Diamond Plasmacytoid Dendritic Cell Isolation Kit II yields PDC populations with excellent purities. The kit involves a two-step procedure, consisting of depletion of non-PDCs and positive selection using MicroBeads against the PDC-specific antigen CD304 (BDCA-4/Neuropilin-1). This kit has been developed based on the first-generation kit (# 130-092-402) for even better performance and a shorter protocol.

Detailed product information

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. This is in contrast to binding of antibodies to the other PDC-specific antigen, CD303 (BDCA‑2).
1, 2

Detailed separation procedure

For the depletion step, all non-PDCs are magnetically labeled by using cocktails containing specific MicroBeads, biotin-conjugated antibodies, and Anti-Biotin MicroBeads. For the subsequent positive selection step, the enriched PDCs are magnetically labeled with CD304 (BDCA-4/Neuropilin-1) Diamond MicroBeads.

Downstream applications

The Diamond Plasmacytoid Dendritic Cell Isolation Kit II was developed for the isolation of PDCs from PBMCs with maximal purity. It is particularly useful if a PDC population with exceptionally high purity is needed as starting material, e.g. for gene expression profiling or protein purification.

Columns

For the first magnetic separation (depletion): LD or autoMACS
®
Columns. For the second magnetic separation (positive selection): MS, LS, or autoMACS Columns.

References for Diamond Plasmacytoid Dendritic Cell Isolation Kit II, human

Publications

  1. Dzionek, A. et al. (2002) Plasmacytoid dendritic cells: from specific surface markers to specific cellular functions. Hum. Immunol. 63: 1133-1148
  2. Colonna, M. et al. (2004) Plasmacytoid dendritic cells in immunity. Nat. Immunol. 5: 1219-1226

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