Table 2.
Antibody sources and methods
| Antigen (antibody type) | Primary Antibody Source | Dilution Used | Colorimetric Immunohistochemical Method | Fluorescent Immunohistochemical Method* | Method Reference for Colorimetric IHC |
|---|---|---|---|---|---|
| TSPO (polyclonal) | Janet Morgan (Roswell Park Cancer Institute) | 1:500 | Avidin-Biotin Complex Method LSAB2 kit (DAKO) with hematoxylin counterstain | Avidin Kit (Vector Laboratories, Ltd., Burlingame, CA) with fluorescein-conjugated streptavidin with DAPI counterstain | [27] |
| TSPO (monoclonal, 8D7) | Pierre Casellas (Sanofi Synthelabo) | 1:350 | ImmPRESS anti-mouse peroxidase kit (Vector Laboratories, Ltd.) with hematoxylin counterstain | Avidin Kit (Vector Laboratories, Ltd.) with fluorescein-conjugated streptavidin with DAPI counterstain | [28] Staining methods were modified for this study. |
| GFAP (polyclonal) | Biogenex (San Ramon, CA) | 1:250 | Primary with goat anti-rabbit alkaline phosphatase-conjugated secondary | Primary with goat anti-rabbit Alexa fluorophor 594-conjugated secondary antibody with DAPI counterstain | [26] |
| CD45RB (monoclonal, clone MT4) | Pharmingen | 1:100 | Not performed for this study | Primary with goat anti-mouse Alexa fluorophor 594-conjugated secondary antibody with DAPI counterstain | [32] |
| CD68 (monoclonal, clone KP1) | DAKO (Carpinteria, CA) | 1:600 | Primary with goat anti-rabbit alkaline phosphatase-conjugated secondary | Primary with goat anti-mouse Alexa fluorophor 594-conjugated secondary antibody with DAPI counterstain | [23] |
| Iba1 (polyclonal) | Wako Pure Chemical Industries, Ltd. (Richmond, VA) | 1:500 | Avidin-Biotin Complex Method with Vectastain kit, Alkaline Phosphatase (Vector Laboratories, Ltd.) | Not performed for this study | [71] |
| HIV-1 p24 (monoclonal, clone MIB-1) | DAKO | 1:10 | Primary with goat anti-rabbit alkaline phosphatase-conjugated secondary | Primary with goat anti-mouse Alexa fluorophor 594-conjugated secondary antibody with DAPI counterstain | [23] |
| SIV mac251 gp41 (monoclonal, clone KK41) | NIH AIDS Research and Reference Reagent Program (Dr. Karen Kent and Miss Caroline Powell) | 1:200 | Avidin-Biotin Complex Method with Vectastain kit, Peroxidase (Vector Laboratories, Ltd.) with hematoxylin counterstain | Not performed for this study | [72] |
| β-amyloid 40 (monoclonal, clone 11A5) | Peter Davies (Albert Einstein College of Medicine) | 1:1000 | Avidin-Biotin Complex Method with Vectastain kit, Peroxidase (Vector Laboratories, Ltd.) | Not performed for this study | P. Davies, personal communication |
*
Fluorescent methods were devised for the current study.