YILIMART - Plasmacytoid Dendritic Cell Iso Kit, m

The Plasmacytoid Dendritic Cell Isolation Kit is designed for the isolation of untouched mouse plasmacytoid dendritic cells (PDCs) from single-cell suspensions of lymphoid organs.

Background information

Freshly isolated PDCs express low levels of MHC class II and costimulatory molecules and are poor stimulators of T helper cells. Upon stimulation with bacterial DNA containing particular unmethylated CpG motifs (CpG-DNA) or upon viral challenge, they produce large amounts of type I IFN and acquire antigen-presenting capacity.

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PDCs can be directly identified in lymphoid and non-lymphoid organs by the specific expression of mPDCA-1 (murine plasmacytoid dendritic cell antigen-1).

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mPDCA-1 has been identified as bone marrow stromal antigen 2 (BST2).

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Detailed separation procedure

Using the Plasmacytoid Dendritic Cell Isolation Kit, non-PDCs are labeled with a cocktail of biotin-conjugated monoclonal antibodies and Anti-Biotin MicroBeads and depleted by retaining them on a MACS

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Column in the magnetic field of a MACS Separator, while the highly enriched unlabeled PDCs pass through the column.

Downstream applications

PDCs isolated by using the Plasmacytoid Dendritic Cell Isolation Kit can be used to analyze antigen uptake, antigen processing, T cell activation or T cell tolerance induction, cross-priming of cytotoxic T cells, and T helper cell polarization by PDCs.

Columns

MS, LS, or autoMACS

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Columns.

Figure 1

PDCs were isolated from splenocytes by using the Plasmacytoid Dendritic Cell Isolation Kit, a MidiMACS™ Separator, and an LS Column. Cells were fluorescently stained with Anti-mPDCA-1-FITC and Anti-Siglec-H-PE and analyzed by flow cytometry using the MACSQuant

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Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Before separation	Plasmacytoid dendritic cells

Figure 2
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Figure 2

Purity and recovery:
Isolation of plasmacytoid dendritic cells from mouse spleen using the Plasmacytoid Dendritic Cell Isolation Kit, mouse. Percentages of purity and recovery were compared to another commercially available product.

Purity and recovery:
Isolation of plasmacytoid dendritic cells from mouse spleen using the Plasmacytoid Dendritic Cell Isolation Kit, mouse. Percentages of purity and recovery were compared to another commercially available product.
Figure 3
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Figure 3

Residual cell types in target fraction:
Isolation of plasmacytoid dendritic cells from mouse spleen using the Plasmacytoid Dendritic Cell Isolation Kit, mouse. Percentages of other residual cell types in the collected final target fraction were compared to another commercially available product.

Residual cell types in target fraction:
Isolation of plasmacytoid dendritic cells from mouse spleen using the Plasmacytoid Dendritic Cell Isolation Kit, mouse. Percentages of other residual cell types in the collected final target fraction were compared to another commercially available product.

Selected references
1. Blasius, A. L. et al. (2006) Bone marrow stromal cell antigen 2 is a specific marker of type I IFN-producing cells in the naive mouse, but a promiscuous cell surface antigen following IFN stimulation. J. Immunol. 177: 3260-3265
2. Nakano, H. et al. (2001)
  
  CD11c
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  B220
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  Gr-1
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  cells in mouse lymph nodes and spleen display characteristics of plasmacytoid dendritic cells.
  
  J. Exp. Med. 194: 1171-1178
3. Shortman, K. and Liu, Y. J. (2002) Mouse and human dendritic cell subtypes. Nat. Rev. Immunol. 2: 151-161
4. Fischer et al. (2004) MACS&more 8(1): 13-15

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Plasmacytoid Dendritic Cell Iso Kit, m

For research use only

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Background information

Detailed separation procedure

Downstream applications

Columns

Figure 1

Figure 2

Figure 2

Figure 3

Figure 3

Selected references

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