The CD8
+
Dendritic Cell Isolation Kit is an indirect magnetic labeling system for the positive selection of CD8
+
DCs from single-cell suspensions of spleen and lymph node.

Data and images for
CD8
+
Dendritic Cell Isolation Kit
, mouse

Figures

Figure 1

Isolation of CD8
+
DCs from a mouse spleen cell suspension using the CD8
+
Dendritic Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator.
Spleen cells before separation
Pre-enriched CD8
+
CD11c
+
DCs
View details

Figure 1

Isolation of CD8
+
DCs from a mouse spleen cell suspension using the CD8
+
Dendritic Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator.
View details

Figure 1

Isolation of CD8
+
DCs from a mouse spleen cell suspension using the CD8
+
Dendritic Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator.
Isolated CD8
+
CD11c
+
DCs
View details

Figure 1

Isolation of CD8
+
DCs from a mouse spleen cell suspension using the CD8
+
Dendritic Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator.

Figure 2

View details
Working scheme of the CD8
+
Dendritic Cell Isolation Kit.

Figure 2

Working scheme of the CD8
+
Dendritic Cell Isolation Kit.

Specifications for
CD8
+
Dendritic Cell Isolation Kit
, mouse

Overview

The CD8
+
Dendritic Cell Isolation Kit is an indirect magnetic labeling system for the positive selection of CD8
+
DCs from single-cell suspensions of spleen and lymph node.

Detailed product information

Background information

CD8
+
DCs coexpress CD11c, CD205, MHC class II, CD40, CD80, CD86, and are negative for CD4 and CD11b. In spleen and lymph nodes, CD8
+
DCs are found at moderate levels (23% and 17% of all CD11c
+
DCs, respectively) and are located in the T cell areas. Skin-draining lymph nodes harbor an additional CD8
+
DC subset (CD4
-
CD8
+
CD11b
+
CD205
+
) which accounts for 33% of all CD11c
+
DCs in this organ and is thought to represent the mature form of Langerhans cells.

Detailed separation procedure

The isolation of CD8
+
DCs is performed in two steps. First, DCs are enriched by depletion of T cells, NK cells, and B cells with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. Subsequently, CD8
+
DCs are positively selected from the enriched DC fraction with CD8a MicroBeads.

Downstream applications

CD8
+
DCs isolated using the CD8
+
Dendritic Cell Isolation Kit are suitable to analyze antigen uptake, antigen processing, T cell activation or tolerance induction, cross-priming of cytotoxic T cells, and T cell polarization by CD8
+
DCs. Isolated splenic CD8
+
DCs have been analyzed for the expression of a signal regulatory protein, which plays a role in the migration of murine Langerhans cells.
1
They additionally have been isolated and adoptively transferred to study their role in the induction of regulatory CD4
+
T cells.
2
The CD8
+
Dendritic Cell Isolation Kit was also used to analyze the expression and regulation of EBV-Induced Gene 3 (EBI3) in mouse DC subsets.
3

Columns

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

References for
CD8
+
Dendritic Cell Isolation Kit
, mouse

Publications

  1. Hagnerud, S. et al. (2006) Deficit of CD47 results in a defect of marginal zone dendritic cells, blunted immune response to particulate antigen and impairment of skin dendritic cell migration. J. Immunol. 176: 5772-5778
  2. Stock, P. et al. (2004) Induction of T helper type 1-like regulatory cells that express Foxp3 and protect against airway hyper-reactivity. Nat. Immunol. 5: 1149-1156
  3. Wirtz, S. et al. (2005) EBV-induced gene 3 transcription is induced by TLR signaling in primary dendritic cells via NF-kappa B activation. J. Immunol. 174: 2814-2824

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+
Dendritic Cell Isolation Kit
, mouse

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