YILIMART - CD56 CD16- NK Cell Isolation Kit, human

The CD56

⁺

CD16

^–

NK Isolation Kit was developed for the isolation of CD56

^bright

CD16

^–

NK cells from PBMCs by sequential sorting. CD56

^bright

CD16

^–

NK cells have a high capacity for cytokine production and are thus suggested to have an immunoregulatory role.

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.

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:	B:
PBMCs before separation	Pre-enriched CD56 ⁺ CD16 ^– NK cells after depletion of non-NK cells and CD56 ⁺ CD16 ⁺ NK cells

C:
Isolated CD56 ⁺ CD16 ^– NK cells

Selected references
1. Deniz, G. et al. (2002) Human NK1 and NK2 subsets determined by purification of IFN-gamma-secreting and IFN-gamma-nonsecreting NK cells. Eur. J. Immunol. 32: 879-884
2. Gerosa, F. et al. (2005) The reciprocal interaction of NK cells with plasmacytoid or myeloid dendritic cells profoundly affects innate resistance functions. J. Immunol. 174: 727-734
3. Romagnani, C. et al. (2005)
  
  Activation of human NK cells by plasmacytoid dendritic cells and its modulation by CD4
  ⁺
  T helper cells and CD4
  ⁺
  CD25
  ^hi
  T regulatory cells.
  
  Eur. J. Immunol. 35: 2452-2458
4. Hagberg, N. et al. (2011) IFN-α production by plasmacytoid dendritic cells stimulated with RNA-containing immune complexes is promoted by NK cells via MIP-1β and LFA-1. J. Immunol. 186: 5085-5094
Brochures and posters

Natural killer cell research

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CD56 CD16- NK Cell Isolation Kit, human

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Background information

Detailed separation procedure

Downstream applications

Columns

Figure 1

Selected references

Brochures and posters

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