UNIT 3.22 Isolation of Murine Natural Killer Cells

  1. Melissa A. Pak-Wittel1,
  2. Sytse J. Piersma1,
  3. Beatrice F. Plougastel1,
  4. Jennifer Poursine-Laurent1,
  5. Wayne M. Yokoyama1,2

Published Online: 2 APR 2014

DOI: 10.1002/0471142735.im0322s105

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Pak-Wittel, M. A., Piersma, S. J., Plougastel, B. F., Poursine-Laurent, J. and Yokoyama, W. M. 2014. Isolation of Murine Natural Killer Cells. Current Protocols in Immunology. 105:IV:3.22:3.22.1–3.22.9.

Author Information

  1. 1

    Division of Rheumatology, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri

  2. 2

    Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri

Publication History

  1. Published Online: 2 APR 2014


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. Curr. Protoc. Immunol.. 105:3.22.1-3.22.9. © 2014 by John Wiley & Sons, Inc.


  • immunology;
  • innate immunity;
  • cell isolation;
  • magnetic separation