Cd11b antibody

Neuronal expression was assessed using a neuron-specific marker, NeuN antibody (1∶500 dilution, Chemicon), which is a DNA binding protein that will bind to the nucleus of neurons. Activated microglia and brain macrophages were observed using the CD11b antibody (1∶1000 dilution, Serotec) which is commonly known as OX42 and used to visualise activated microglia [21] (link), [22] . Frozen coronal cryostat sections (16 µm) were post-fixed in paraformaldehyde (4%). Slides were incubated overnight with either the NeuN or OX42 antibody at 4°C. Sections were then incubated at room temperature with a fluorescence labelled secondary antibody, Alexa 488 (1∶500 dilution, Invitrogen). The number of immuno-positive cells were counted within a 1 mm² site for the NeuN antibody and a 0.5 mm² site for the OX42-immuno staining in the infarcted region of the ipsilateral hemisphere. Ballistic light images from consecutive sections for each individual rat were used to locate the infarct and non-infarct regions in which cell counts were conducted. As a control, the number of immuno-positive cells in the identical region in the contralateral hemisphere was also counted. In addition, the number of immuno-positive cells in within the non-infarcted region of the ipsilateral hemisphere and matched region on the contralateral hemisphere was also calculated.

N-Desmethylclozapine, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), MPP⁺, and Leu methyl ester (LME) were purchased from Sigma-Aldrich (St. Louis, MO). Clozapine and clozapine N-oxide were obtained from the NIMH Chemical Synthesis and Drug Supply Program (Research Triangle Institute, RTP NC). Lipopolysaccharide (LPS strain O111:B4) was purchased from Calbiochem (San Diego, CA). WST-1 was purchased from Dojindo Laboratories (Gaithersburg, MD). Cell culture ingredients were obtained from Invitrogen (Carlsbad, CA). [³H]DA was purchased from PerkinElmer Life Sciences (Boston, MA). The polyclonal anti-tyrosine hydroxylase (TH) antibody was purchased from CHEMICON International (Temecula, CA). The polyclonal ionized calcium-binding adaptor molecule 1 (Iba-1) antibody was purchased from Wako Chemicals USA (Richmond, VA). The monoclonal CD11b antibody was purchased from AbDSerotec (Raleigh, NC). The biotinylated secondary antibodies were purchased from Vector Laboratories (Burlingame, CA).