PAD4 detected by Flow Cytometry
50μL of peripheral blood was lysed with 1mL of FACS Lysing Solution (555899, BD, USA) in dark for 10 minutes. CD16 (563830, BD, USA) and CD62L (555543, BD, USA) were used for neutrophil staining. Then, after fixing and permeabilizing, cells were incubated with mouse anti-human PAD4 as primary antibody (ab128086, Abcam, UK) for 1 hour, and stained with PE-rat anti-mouse IgG2a as secondary antibody (12-4210-82, Thermo Fisher, USA) for 30 minutes. Data were detected by a flow cytometer (FACS Calibur, BD), and analyzed with FlowJo7.6 software.