NK1.1+ iNKT Cell Isolation Kit ( #130-096-513 )

NK1.1⁺iNKT Cell Isolation Kit

Description

The NK1.1⁺ iNKT Cell Isolation Kit was developed for the isolation of NK1.1-expressing NKT cells from murine lymphoid tissues by sequential sorting.

Details

Background information
Natural killer T (NKT) cells are a heterogeneous group of lymphocytes that share properties of both T cells and natural killer (NK) cells. They are an important link between the innate and the adaptive immune system, promoting or suppressing immune responses.
In C57BL/6 mice, NKT cells were first defined according to their expression of the NK cell-associated marker NK1.1 (CD161). The term iNKT cells preferentially refers to CD1d-restricted T cells, expressing a heavily biased, semiinvariant T cell receptor (TCR Vα14-Jα18, Vs8.2, Vs7 and Vs2).

Detailed separation procedure
The isolation of NK1.1⁺ NKT cells is performed in a two-step procedure. First, non-target cells like NK cells, B cells, Macrophages, CD8⁺ T cells and TCRγδ⁺ T cells are magnetically labeled and depleted using a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. Subsequently, NK1.1⁺ iNKT cells are positively selected from the pre-enriched fraction using Anti-NK1.1-APC and Anti-APC MicroBeads.

Columns

For the first magnetic separation (depletion): LD or autoMACS Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns

Further information

Isolation of iNKT cells from mouse spleen using CD1d tetramers
[PDF; 252,4 KB]

Figure 1

Splenocytes from a C57BL/6 mouse were isolated by using the NK1.1⁺ iNKT Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator. Cells were fluorescently stained with Anti-NK1.1-APC, CD3ε-VioBlue, and α-Gal-Cer/CD1d Tetramer-PE and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Before separation

After separation