The CD3+CD56+ NKT Cell Isolation Kit was developed for the sequential separation of CD3+CD56+ natural killer (NK) T cells from human PBMCs and other single-cell suspensions.
NKT cells represent a subpopulation of T cells that possesses properties of NK cells. NKT cells can be stimulated through contact with antigen or by cytokines such as IL-12 to release large amounts of cytokines and to exert cytotoxic effects. NKT cells are a crucial part of the innate immune system and are involved in the development of autoimmune diseases, in tumor immunology as well as in immunity against viruses, bacterial, fungal, and parasitic pathogens.
Detailed separation procedure
In a first step, NK cells and monocytes are magnetically labeled by using a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted by separation over a MACS® Column. In the second step, CD3+CD56+ NKT cells are isolated by positive selection from the pre-enriched NKT cell fraction using CD56 MicroBeads.
CD3+CD56+ NKT cells can be used for a variety of studies such as analysis of cytokine secretion, gene expression analysis of cell subsets, analysis of the functional role of cell surface receptors, cytotoxic and cytolytic activity.
For the first magnetic separation (depletion): LD or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.
CD3+CD56+ NKT cells were isolated from human PBMCs by using the CD3+CD56+ NKT Cell Isolation Kit, an LD Column and an MS Column, a MidiMACS™ Separator and a MiniMACS™ Separator. The cells were fluorescently stained with CD3-FITC and CD56-APC.
Enriched CD3+CD56+ NKT cells