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Published on: October 14, 2016
Protocol for multi-organ isolation of murine leukocytes for flow cytometry
Paul Jerard Layug1, Agata Marcinow2, David Sontag3
1Department of Internal Medicine, University of Manitoba, Winnipeg, MB, Canada; Manitoba Centre for Proteomics and Systems Biology, Winnipeg, MB, Canada.
The immune system comprises several specialized cells across multiple organs. Here, we present a protocol for the isolation of leukocytes from murine blood, spleen, mesenteric lymph nodes, liver, kidney, and skeletal muscle. We describe steps for cell isolation and gradient density separation and provide four antibody panels to identify adaptive immune cells (e.g., CD4+ and CD8+ T cell subsets) by flow cytometry. This protocol provides a basic framework for phenotyping immune cells from in vivo mouse models.
The immune system comprises several specialized cells across multiple organs. Here, we present a protocol for the isolation of leukocytes from murine blood, spleen, mesenteric lymph nodes, liver, kidney, and skeletal muscle. We describe steps for cell isolation and gradient density separation and provide four antibody panels to identify adaptive immune cells (e.g., CD4+ and CD8+ T cell subsets) by flow cytometry. This protocol provides a basic framework for phenotyping immune cells from in vivo mouse models.
