Aβ1 42

Animals were deeply anesthetized with isoflurane, and then perfused with 4% paraformaldehyde in 0.1 M phosphate buffer. Brains were sucrose protected and coronal sections cut on a cryostat (50- μm thick). After blocking nonspecific sites with a buffer containing 2% goat serum, 1% bovine serum albumin (BSA), and 0.2% Triton X-100 in PBS, sections were incubated with primary antibody. The primary antibodies were diluted at 1:500 unless otherwise stated. Antibodies included mouse monoclonals against: MBP (Millipore, 1:10 dilution); N-Sphingomyelinase2 (Abcam); Myelin-associated glycoprotein (MAG, Abbiotec); Lamp1 (Santa Cruz); Caveolin 1 (Cell Signaling); rodent Aβ (Covance); Aβ_1 - 40/42 (Millipore); Aβ_1 - 42 (Covance); and a rabbit polyclonal against neurofilament light chain protein (NF-L) (Millipore; 1:1000 dilutions). The primary antibody was incubated with sections at 4°C overnight. The secondary antibody was incubated with sections at room temperature for 2 h. Goat anti-mouse or goat anti-rabbit Alexa Fluor ^® 488 or 594 conjugated antibodies (Invitrogen) were used for secondary antibodies depending on the species of the primary antibody. Primary antibody was omitted to assess non-specific staining. Sections were stained with FSB, a Congo red derivative [21 (link)] (Millipore, 0.5 μM in PBS, 20 min), to identify amyloid plaques in the mouse AD model.