Macsxpress neutrophil isolation

Blood was obtained from study participants after informed consent through a protocol approved by the University of Wisconsin Internal Review Board (IRB) in accordance with relevant guidelines and regulations, including the Policy for the Protection of Human Subjects. Peripheral blood neutrophils were purified by negative antibody selection using the MACSxpress Neutrophil Isolation and MACSxpress Erythrocyte Depletion kits (Miltenyi Biotec Inc., Auburn, CA)^{12 (link)}. Neutrophils were enumerated by hemocytometer and resuspended in RPMI 1640 (without phenol red) supplemented with 2% heat-inactivated fetal bovine serum (FBS) and supplemented with glutamine (0.3 mg/ml) for all experiments. Incubations were at 37 °C with 5% CO₂.

Blood was obtained from donors who provided written informed consent through a protocol approved by the University of Wisconsin Internal Review Board. Negative antibody selection via the MACSxpress Neutrophil Isolation and MACSxpress Erythrocyte Depletion Kits (Miltenyi Biotec Inc., Auburn, CA, USA) was performed to purify primary human neutrophils. Neutrophils were resuspended in RPMI 1640 (no phenol red) supplemented with 2% FBS. For all experiments incubations were performed at 37 °C with 5% CO₂.