Studied mouse line J20 having (hAPP SweInd) FAD mutations and line I5 wildtype hAPP (hAPPWT) and C57BL/6J non-TG breeders.
Immunofluorescent double staining or general avidin-biotin/peroxidase staining was performed on human brains obtained from Alzheimer's Disease Research Center with the following antibodies: rabbit anti-calbindin (1:15,000, Swant, Bellinzona, Switzerland); rabbit anti-c-Fos (1:10,000, Ab-5, Oncogene); mouse monoclonal anti-Neu-N (1:5,000, Chemicon); or mouse monoclonal anti-Abeta 3D6
(1:500, Elan Pharmaceuticals, South San Francisco, CA), and then with fluorescein-labeled donkey anti-rabbit (1:300, Jackson ImmunoResearch); Texas red-labeled donkey anti-mouse (1:300, Jackson ImmunoResearch); biotinylated goat anti-rabbit (1:200, Vector Laboratories); or biotinylated goat anti-mouse (1:600, Vector Laboratories). Diaminobenzidine was used as a chromagen. Immunofluorescence was visualized by confocal microscopy.
Western Blots were probed with rabbit anti-CB (1:20,000, Swant); mouse monoclonal anti-hApp 8E5 (1:1,000, Elan Pharmaceuticals); or mouse monoclonal anti-alpha-tubulin B512 (1:100,000, Sigma).