Anti cd11b antibody

Mouse eyes were dissected and fixed in paraformaldehyde before embedding in paraffin, sectioning at 10-μm thickness and staining with hematoxylin and eosin. For immunohistochemistry staining, the sections were de-paraffinated and rehydrated followed by incubated with citrate buffer for antigen retrieval. To block endogenous peroxidase activity, the sections were treated with 3% H₂O₂ for 25 minutes. The sections were then blocked with 3% BSA at room temperature for 30 minutes, followed by incubating with anti-CD4+ (1:100, Servicebio, cat: gb13064), anti-CD8+ (1:1000, Servicebio, cat: gb11068), anti-PD-L1 antibody (1 ug/mL, Abcam, cat: 238697), anti-CD11b antibody (1:500, Servicebio, cat: gb11058), and anti-F4/80 antibody (1:500, Servicebio, cat: gb11027), respectively, at 4°C overnight. The slides were then incubated with an anti-rabbit secondary antibody (1:500, Servicebio, cat: gb23303) or anti-mouse secondary antibody (1:500, Servicebio, cat: gb23301), followed by incubating with the freshly prepared DAB substrate solution to reveal the color of antibody. At last, the tissue slides were counterstained with hematoxylin and blued with ammonia water, and then dehydrated and coverslipped. Images were collected by a fluorescence microscope (Eclipse Ni, Nikon).