Microbeads
CD8+ Dendritic Cell Isolation Kit , mouse
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.
Quantity
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.
3Columns
For the first magnetic separation (depletion): LD or autoMACS
®
Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.