Goat anti mouse cd31 polyclonal antibody

In the distribution study of antibodies, the mice were injected with ~300 µg of 1849-whole IgG-AF647 and 1849-Fab-AF647 via the tail vein. The mice were euthanized under deep anesthesia at 3 h after the injection, and the tumors were excised. The tumors were then embedded in Tissue-Tec optimal-cutting-temperature compound (Sakura Finetek, Tokyo, Japan) and frozen at -80˚C until use. Frozen sections, 6-µm-think, of the tumors were fixed with 4% paraformaldehyde in PBS for 15 min at RT and blocked with 5% skim milk in PBS for 1 h at RT. The sections were then incubated with goat anti-mouse CD31 polyclonal antibody (2 µg/ml; R&D Systems, Minneapolis, MN, USA) for 1 h at RT, and then with AF555 conjugated anti-goat polyclonal antibody (4 µg/ml; Invitrogen) for 1 h at RT. Subsequently, the sections were stained with DAPI solution (1 µg/ml) for 5 min at RT and the slides were covered with Fluoromount-G. Fluorescence images were obtained with BIOREVO BZ9000.