Isolation of murine natural killer cells.
This unit describes the isolation of natural killer (NK) cells from mouse spleen. The basic protocol describes a method for preparing a highly purified NK cell population from mouse spleen by depletion of contaminating cells with selected monoclonal antibodies (MAbs) and magnetic separation. There are several advantages to this negative selection process. One of these is that the NK cells are not coated with antibody and, therefore, are not at risk of functional perturbation by antibody cross-linking. Additionally, negative selection provides a way to isolate diverse subpopulations of NK cells without selectively purifying a specific subpopulation. Following enrichment, NK cell purity can be assessed by cell surface phenotype using flow cytometry.
Division of Rheumatology, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri.
Melissa A
MA
Pak-Wittel
Division of Rheumatology, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri.
Sytse J
SJ
Piersma
Division of Rheumatology, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri.
Beatrice F
BF
Plougastel
Division of Rheumatology, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri.
Jennifer
J
Poursine-Laurent
Division of Rheumatology, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri.|Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri.
Wayne M
WM
Yokoyama
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology; D051379:Mice; D013154:Spleen / Q000166:cytology / Q000276:immunology
D000911:Antibodies, Monoclonal
cell isolation; immunology; innate immunity; magnetic separation