Cd11c ep1347y

Paraffin-embedded full thickness placental samples were sliced into 3 µm sections, dewaxed and heated in 0.01 M pH 6 sodium citrate buffer for antigen retrieval. After blocking endogenous peroxidase activity and non-specific binding, the sections were incubated overnight with the following primary antihuman antibodies: mouse monoclonal anti-CD3 (F7.2.38, 1:70, Dako, Glostrup, Denmark), mouse monoclonal anti-CD20 (L26, 1:126, Dako), mouse monoclonal anti-CD68 (PG-M1, 1:30, Dako), rabbit monoclonal anti-CD11c (EP1347Y, 1:500, Abcam, Cambridge, UK) and rabbit polyclonal anti-ACKR2 (1:400, Sigma-Aldrich, St Louis, MO, USA). Sections were then incubated with the appropriate chromogenic secondary antibody (ImmPRESS Polymer Detection Kit, anti-rabbit and anti-mouse, Vector Laboratories, Burlingame, CA, USA). The immunoreactivity was developed using 3,3′-diaminobenzidine tetrahydrochloride (Vector Laboratories) as chromogen. Isotype-matched control antibodies were included as a negative control and tonsil sections as a positive control. The sections were observed under a light microscope (Olympus BX43, Olympus, Tokyo, Japan) and photographed by digital camera (DP22 using Olympus Cell Sense Entry 2.2 for imaging acquisition).