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.

Data and images for
NK1.1
+
iNKT Cell Isolation Kit
, mouse

Figures

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.
Unseparated fraction
A:
B:
View details

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.
View details

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.
After separation
A:
B:
View details

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.
View details

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.

Specifications for
NK1.1
+
iNKT Cell Isolation Kit
, mouse

Overview

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.

Detailed product information

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.

Resources for
NK1.1
+
iNKT Cell Isolation Kit
, mouse

References for
NK1.1
+
iNKT Cell Isolation Kit
, mouse

Publications

  1. Godfrey, D. I. et al. (2004) NKT cells: what's in a name? Nat. Rev. Immunol. 4(3): 231-237
  2. Bendelac, A. et al. (2007) The biology of NKT cells. Annu. Rev. Immunol. 25: 297-336
  3. Godfrey, D. I. et al. (2010) Raising the NKT cell family. Nat. Immunol. 11(3): 197-206

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+
iNKT Cell Isolation Kit
, mouse

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