Monoclonal rat anti mouse cd11b antibody

Our phagocytic model used N9 cells to engulf fluorescent bioparticles (see detailed description below). Round glass cover slips were placed in the well of 24-well plates before seeding N9 cells. After the 20 min EMF exposure, bioparticles (5 × 10⁶ per well) were added to all wells and incubated for different times. Bioparticles were added 1 h before the indicated time points that the cells were tested in the phagocytosis assay. Then the glass cover slips were carefully taken out of the wells at different times and rinsed twice in PBS. After being fixed and permeabilized, the cells were blocked with goat serum (ZsBio) for 20 min at room temperature and washed three times in PBS. The cells were incubated with the rat anti-mouse monoclonal antibody CD11b (1:100) (AbD Serotec) at 37°C for 1 h. After washing three times in PBS, the cells were incubated with the goat anti-rat Alexa Fluor® 488 secondary antibody (1:200) (Molecular Probes, OR, USA) for 1 h at 37°C in the dark. The cover slips with cells were washed three times in PBS and mounted with an aqueous-based anti-fade mounting medium on glass slides. Images of stained cells were captured using a Leica TCS-SP5 confocal laser scanning microscope (Leica, Mannheim, Germany). The confocal images were acquired using Imaris software (version 7.6; Bitplane, Zurich, Switzerland).