Anti cd163 ab182422

Penicillin, fetal bovine serum (FBS), DMEM/ F12 medium, and PBS were purchased from GE™ Hyclone. APS was purchased from Beijing Solarbio Science & Technology Co., Ltd. ATP was purchased from Shanghai Zeye Biological Technology Co., Ltd. Rat ATP ELISA Kit and rat ADP ELISA Kit were purchased from Beijing winter song Boye Biotechnology Co. Ltd. EBD was purchased from Real-Times (Beijing) Biotechnology Co., Ltd. EDTA antigen repair solution was purchased from Servicebio. Antibodies: anti-GAPDH (KC-5G5, KangChen Bio-tech), anti-P2X7R (BA2808, Boster Biological Technology Co., Ltd.), anti-CD163 (ab182422, Abcam), anti-CD206 (sc-58,986, Santa Cruz), anti-CD86 (NBP2-25,208, Novus), anti-HLA-DR (MA5-32,232, Invitrogen), anti-CD163-PE (85–12-1639-41, eBioscience), anti-CD206-APC (85–17-2069-41, eBioscience), anti-CD86-PE (70-AR08604-100, MultiSciences), and anti-HLA-DR-APC (70-AH0HD05-20, MultiSciences).

The mIHC was performed using a PE‐Opal 7‐color Automation IHC Kit (NEL821001KT, Perkin Elmer). FFPE blocks were heated at 65°C for 30 min. The slides were then dewaxed, rehydrated, and fixed using a Leica BOND RX auto Stainer (Leica Biosystems). Subsequently, the slides were stained with three markers: anti‐CD8 (790–4460, Ventana), anti‐CD163 (ab182422, Abcam), and anti‐Pan Keratin (790–2135, VENTANA), followed by incubation with horseradish peroxidase (HRP)‐conjugated secondary antibody and tyramide signal amplification (TSA). Finally, the slides were stained with 4′‐6′‐diamidino‐2‐phenylindole (DAPI) for 10 min at 1:10 dilution, and images were acquired using a Vectra 3.0 pathology imaging system microscope (PerkinElmer Inc.).