Description
Background information
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 minority of all NK cells in the blood. They lack expression of killer cell immunoglobulin-like receptors (KIRs) and, accordingly, neither show strong cell-mediated cytotoxicity nor antibody-dependent cell-mediated cytotoxicity (ADCC). They are potent producers of cyokines, predominantly IFN-γ, thus linking innate and adaptive immune system.1-3
Detailed separation procedure
The isolation of CD56+CD16- NK cells is performed in a two-step procedure. First, non-NK cells and CD56+CD16+ NK cells are labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads and 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 CD56 MicroBeads and isolated by positive selection from the pre-enriched cell fraction by using a second column.
Downstream applications
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 interactions.4 Furthermore, cells can be used to study the role of NK cells in diseases or pregnancy.
Columns
For the first magnetic separation (depletion): LS or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.
Overview
The CD56+CD16– NK Isolation Kit was developed for the isolation of CD56brightCD16– NK cells from PBMCs by sequential sorting. CD56brightCD16– NK cells have a high capacity for cytokine production and are thus suggested to have an immunoregulatory role.
