Cy3 conjugated goat anti rabbit immunoglobulin g igg antibody

Human blood neutrophils were pretreated with or without PD for 1 h. These prepared neutrophils were placed on cytospins and stimulated with 25 nM PMA or PBS for 4 h. Alternatively, bone marrow-derived neutrophils from PD-, solvent-, or saline-treated mice were directly placed on cytospins and cultured in the absence of serum for 4 h. And then all those neutrophils were fixed with 1 ml PBS/1% paraformaldehyde. After blocking with 5% fetal bovine serum, the cytospins were stained with rabbit anti-myeloperoxidase (MPO) antibody (Abcam, catalog ab208670, Cambridge, UK) in combination with a second Cy3-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (Servicebio, catalog GB21303, Boston, MA, USA). The DNA was visualized by 4′, 6′-diamidino-2-phenylindole (DAPI) (Thermo Fisher Scientific). After being mounted, specimens were analyzed using a fluorescence microscope.