Anti mouse hq

Sections of 4µm of FFPE samples were used for immunohistochemistry analysis. To visualize M1 and M2 macrophage subsets in placental tissue, the chromogenic duplex staining was done with CD68 and CD163 by an automated staining procedure using the Ventana Benchmark Discovery (Ventana Medical Systems Inc.). In brief, following deparaffinization and heat-induced antigen retrieval with CC1 (#950-500, Ventana) for 40 minutes at 95°C, the tissue samples were firstly incubated with CD68 for 60 minutes at 37˚C, followed by detection with anti-mouse HQ (#760-4814, Ventana) for 16 minutes and subsequently anti-HQ HRP (#760-4820), and visualized with Discovery Purple (#760-229, Ventana) for 32 minutes. An antibody denature step was performed using CC2 (#950-123, Ventana) for 20 minutes at 100˚C. Secondly, CD163 was incubated for 32 minutes at 37˚C, followed by detection with anti-mouse HQ and subsequently anti-HQ HRP, and visualized with Discovery Teal (#760-247, Ventana) for 32 minutes. Finally, all slides were counterstained with hematoxylin II (#760-2208) and bluing reagents (#760-2037) for 4 minutes. Antibody information and clonality can be found in Supplementary Table S3.