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. 2017 Jul 5;7(13):e2386. doi: 10.21769/BioProtoc.2386

Figure 1. Neutrophil isolation.

Figure 1.

A. Human neutrophils were isolated from fresh peripheral blood of healthy donors by gradient centrifugation (step A1). The neutrophil layer at the interface of H1077 and H1119 were collected. PBMC, peripheral blood mononuclear cells; PMN, polymorphonuclear leukocytes; RBC, red blood cells. B. The cell viability was analyzed with PI dye and the purity of the viable neutrophils was analyzed using a human CD15 antibody within the PI negative gate. C. Human neutrophils were isolated from buffy coat with the MACSxpress method (step A2). The cell viability was analyzed with PI dye and the purity of the viable neutrophils was analyzed with human CD15 antibody within the PI negative gate. D. Murine bone marrow neutrophils were isolated by magnetic sorting using a negative depletion method (step A3). The cell viability was analyzed with PI dye and the purity of the viable neutrophils was analyzed with a mouse Ly6G (Gr-1) antibody within the PI negative gate. Gray histograms: PI dye or CD15 antibody or Gr-1 antibody; open histogram: unstained control or isotype antibody.