Rat anti cd200

Animals were anesthetized with a lethal dose of pentobarbital and brains were perfused via the ascending aorta with ice-cold PBS followed by cold 4% paraformaldehyde. Tissues were deep frozen in dry ice, serial 10 μM coronal sections were prepared and immunofluorescent stainings were performed as previously described (Einstein et al., 2003 (link)). The following antibodies were used: rabbit anti-glial fibrillary acidic protein (1:200, GFAP, Dako), rat ant-BrdU (1:200, Serotec), rabbit anti-Iba1 (1:250, Wako), rat anti-CD3 (1:200, AbD Serotec) and rat anti-CD200 (1:100, Abcam). Goat anti rat Alexa-fluor555 (1:200), goat anti rabbit Alexa-fluor555 (1:200) and goat anti rabbit Alexa-fluor488 (1:200) were used as secondary antibodies where appropriate. When staining sections containing GFP+ cell grafts, Alexa-fluor555 conjugated secondary antibodies were used. For in vitro staining, floating spheres or single cell astrocytes were adhered to 35-mm tissue culture dishes coated with 10 μg/ml poly-D-lysine and 5 μg/ml fibronectin (Sigma). In vitro staining was performed as previously described (Einstein et al., 2006a (link)).