Rat anti cd11b

The protocol for immunohistochemical staining has been described elsewhere35 (link). The 4% paraformaldehyde-fixed brain sections were stained using rabbit anti-ionized calcium-binding adapter molecule-1 (Iba1) (1:2000) (Wako Pure Chemical Industries, Osaka, Japan) and rat anti-CD11b (1:200) (R&D Systems, Minneapolis, MN) for microglia, mouse anti-major histocompatibility complex (MHC) class II (1.:250) (AbD Serotec, Oxford, UK) for activated microglia and rabbit anti-tyrosine hydroxylase (TH) (1:2000) (Chemicon, Temecula, CA) for DA neurons. Brain specimens were incubated with appropriate biotin-conjugated secondary antibodies and avidin-biotin peroxidase (ABC; Vector Laboratories, Burlingame, CA) using diaminobenzidine as the substrate. The signals were evaluated by the morphologies and intensities after subtracting the signals of the primary antibody-omitted negative controls. In some cases, the primary antibodies were replaced by isotype antibodies (normal anti-rabbit, anti-rat and anti-mouse antibodies) to control for non-specific binding of the primary antibodies. Incubations without primary antibody were used as negative controls.