| Description |
NK cells are not a homogeneous cell population and can be divided into several subsets according to functional and phenotypic differences. The CD56+CD16+ NK Isolation Kit was developed to isolate an NK cell population that is CD56dim and CD16+. Representing about 7% (range: 2–14%) in PBMCs, CD56+CD16+ cells are the major NK cell subset in blood. These cells also express killer cell immunoglobulin-like receptors (KIRs) and CD94-associated lectin-like NKG2 receptors. Accordingly, they exhibit strong antibody-dependent cell-mediated cytotoxicity (ADCC) and natural cell-mediated cytotoxicity1. Furthermore, CD56+CD16+ NK cells have a granular phenotype. Unlike CD56+CD16– NK cells, they have a low cytokine production capacity. Gene expression profiling on freshly isolated CD56+CD16– NK cells versus CD56+CD16+ NK cells has revealed several differentially expressed genes.
The isolation of CD56+CD16+ NK cells is performed in a two-step procedure. First, non-NK cells (i.e. T cells, B cells, dendritic cells, stem cells, monocytes, granulocytes, and erythroid cells) are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies against lineage-specific antigens and a cocktail of MicroBeads. Upon subsequent magnetic separation of the cells over a MACS® Column that is placed in a magnetic field of a MACS Separator, the magnetically labeled non-NK cells are retained within the column while the unlabeled NK cells run through. In the second step, the pre-enriched NK cells are directly labeled with CD16 MicroBeads. Upon subsequent magnetic separation, the CD56+CD16+ NK cells are eluted after removing the column from the magnetic field. |
| Applications |
| The CD56+CD16+ NK Isolation Kit was developed for specific isolation of CD56+CD16+ NK cells from PBMCs, for example, for phenotypical and functional characterization of NK cell subsets, for the analysis of receptor expression, cytokine secretion, or cell-cell interaction with other cells of the innate and adaptive immune system. Isolated NK cells can also be used to study the role of NK cells in diseases, e.g., hepatitis infection or cancer, as well as during pregnancy. |
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| Figure 1 |
| CD56+CD16+ NK cells were isolated from human PBMCs using the CD56+CD16+ NK Cell Isolation Kit, an LS and an MS Column, a MidiMACS™ and a MiniMACS™ Separator. Cells were fluorescently stained with CD16-FITC and CD56-PE. |
| A: PBMCs before magnetic separation |
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| B: Enriched NK cells after depletion of non-NK cells |
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| C: Isolated CD56+CD16+ NK cells |
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