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.