The CD56+CD16+ NK Isolation Kit was developed for the isolation of CD56dimCD16+ NK cells from PBMCs by sequential sorting. The CD56dim NK cell subset is highly cytotoxic even without stimulation, but has little to no cytokine production.
NK cells represent a highly specialized lymphocyte population of the innate immune system playing an important role in early immune response.
NK cells can be divided into several subsets according to function and phenotype. CD56+CD16+ NK cells represent the majority of all NK cells in the blood. They express killer immunoglobulin-like receptors (KIRs) and CD16 and, accordingly, show strong cell-mediated cytotoxicity as well as antibody-dependent cell-mediated cytotoxicity (ADCC). Unlike CD56+CD16- NK cells, they have a low cytokine production capacity.1
Detailed separation procedure
The isolation of CD56+CD16+ NK cells is performed in a two-step procedure. First, non-NK cells are labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads und subsequently depleted by separation over a MACS® Column, which is placed in the magnetic field of a MACS Separator. In the second step, the target cells are labeled with CD16 MicroBeads and isolated from the pre-enriched cell fraction by positive selection over a second column.
Cells isolated using the CD56+CD16+ NK Cell Isolation Kit can be used for the 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.2 Furthermore, 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.
For the first magnetic separation (depletion): LS or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.
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.
PBMCs before separation
Enriched NK cells after depletion of non-NK cells
Isolated CD56+ CD16+ NK cells