Anti aβ42 antibody

Frozen sections were air-dried at room temperature for 15 min and with 4% paraformaldehyde for 10 min. Then sections were rinsed with PBST (0.25% Triton-X100) and blocked for 30 min at room temperature in 5% goat serum. After incubation with primary antibodies diluted in PBS for 2 h at room temperature, sections were washed with PBS and incubated with secondary antibodies for 1 h at room temperature. Sections were washed again and mounted with mounting medium containing DAPI (Vector Lab, Burlingame, USA). Primary antibodies used were as follows: anti-cleaved caspase-3 (1:300, Cell Signaling Technology, MA, USA), anti-Atg5 (1:400, Santa Cruz, CA, USA), anti-MAP2 (1:400, Santa Cruz, CA, USA), anti-GFAP (1:300, Abcam, Cambridge, UK), and anti-Aβ42 antibody (Invitrogen, CA, USA). All immunofluorescence images were representative of at least three independent animal experiments.

Anti-Aβ₄₂ antibody (Invitrogen 2μg/ml) was used as the coating antibody, and synthetic Aβ₄₂ was used as a standard. The medium samples, as well as the standards, were denatured in 0.5% sodium dodecyl sulfate (SDS) at 90 °C for 5min, followed by a 1:10 dilution (in order to lower the SDS concentration to 0.05% SDS). Biotinylated mAb4G8 (Nordic Biosite, 80070) was used as detection antibody, followed by incubation with SA-HRP (1:2000 Mabtech) for 1h at RT.