Biotin anti mouse ly 6g antibody

RPMI media containing 0.1% FBS was used to flush bone marrow cells from femurs and tibias of C57BL/6 mice (male or female, 6–8 weeks). Following depletion of erythrocytes using Red Blood Cell Lysis Buffer (Sigma, #R7757), cells were re-suspended and incubated sequentially with Biotin anti-mouse Ly6G antibody (BioLegend, #127604) for 20 min and anti-Biotin MicroBeads (Miltenyi Biotec, #130–090-485) for 15 min and then sorted by AutoMACS Cell Separation Instruments (Miltenyi Biotec).
Following the manufacturer's instructions, human neutrophils were extracted from peripheral blood using a kit from Solarbio (#P9040). Briefly, peripheral blood was collected in tube containing anticoagulant, and mixed slowly with reagents A and C, followed by centrifugation at 1000 × g for 20 min. Following centrifugation, the neutrophil-containing layer between reagent A and C was gently removed and lysed with Red Blood Cell Lysis Buffer to remove erythrocytes. Flow cytometry was performed to determine cell purity. Neutrophils with greater than 90% purity were utilized for the following experiments.