Blood Dendritic Cell Isolation Kit II ( #130-091-379 )

Blood Dendritic Cell Isolation Kit II

Description

The Blood Dendritic Cell Isolation Kit II is a magnetic labeling system for the concurrent isolation of plasmacytoid and myeloid dendritic cells from human PBMCs. The isolation is performed in a two-step procedure. First, B cells and monocytes are magnetically labeled and depleted using a cocktail of CD19 and CD14 MicroBeads. Subsequently, the pre-enriched dendritic cells in the non-magnetic flow-through fraction are magnetically labeled and enriched using a cocktail of antibodies against the dendritic cell markers CD304 (BDCA-4/Neuropilin-1), CD141 (BDCA-3), and CD1c (BDCA-1). The highly pure enriched cell fraction comprises of plasmacytoid dendritic cells, CD1c (BDCA-1)⁺ type-1 myeloid dendritic cells (MDC1s), and CD1c (BDCA-1)^- CD141 (BDCA-3)^bright type-2 myeloid dendritic cells (MDC2s). B cells and monocytes are depleted in advance because a subpopulation of B cells expresses CD1c (BDCA-1), and monocytes express CD141 (BDCA-3) at low levels.
The kit includes a Non-Dendritic Cell Depletion Cocktail, a Dendritic Cell Enrichment Cocktail, and FcR Blocking Reagent.
For evaluation of MACS® Separations using the Blood Dendritic Cell Isolation Kit II, staining with the Blood Dendritic Cell Enumeration Kit is recommended, which includes all antibodies necessary for analysis and isotype control as pre-mixed cocktails.

Applications

Dendritic cells isolated by MACS® Technology are, for example, suitable for in vitro induction of primary T cell responses.

Columns

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

Figure 1

Working scheme for the isolation of dendritic cells from PBMCs using the Blood Dendritic Cell Isolation Kit II.

Figure 2

Blood dendritic cells were isolated using the Blood Dendritic Cell Isolation Kit II and stained with the Blood Dendritic Cell Enumeration Kit. Thereby, type-1 myeloid dendritic cells (MDC1s) are identified by expression of CD1c (red dots), type-2 myeloid dendritic cells (MDC2s) by expression of CD141 (blue dots), and plasmacytoid dendritic cells (PDCs) by expression of CD303 (green dots).