Mouse monoclonal antibodies

Synovium was collected by closed Parker-Pearson needle biopsy from one actively inflamed knee joint in RA patient (Schumacher & Kulka, 1972 (link)). As described previously (Ma et al., 2014 (link); Zou et al., 2013 (link)), serial sealed sections of qualified synovial samples were stained with hematoxylin and eosin (H&E) and also immunohistochemically stained with the commercial mouse monoclonal antibodies (Invitrogen, Carlsbad, CA, USA) according to standard staining protocols: anti-CD3 (clone PS1, T cells), anti-CD15 (clone My1, neutrophils), anti-CD20 (clone L26, B cells), anti-CD34 (clone QB End/10, vascular endothelial cells), anti-CD38 (clone SPC32, plasma cells), and anti-CD68 (clone KP1, macrophages). Irrelevant isotype was used as a negative control. Appropriate positive controls were included to rule out the possibility of an absence of staining due to technical failure.

Monocyte/macrophage preparations were routinely analyzed by flow cytometry [Supplementary Figure 1], with mouse monoclonal antibodies from eBioscience. Enriched human peripheral blood monocyte preparations were typically more than 85% CD14⁺CD16⁻ (classical). Upon in vitro differentiation into human macrophages, the cell population was more than 99% CD11b⁺CD14⁺. Mouse monocyte samples were typically more than 80% CD11b⁺Ly-6G⁻. In agreement with previous reports65 (link), enriched mouse peripheral blood monocyte preparations contained a mixture of around 38% Ly-6C^hi and 31% Ly-6C^low subsets, while bone marrow samples contained a higher proportion of around 70% Ly-6C^hi monocytes.