Dylight 488 conjugated goat anti rabbit

Brain samples were fixed with 4% paraformaldehyde for 4 h, followed by overnight immersion in phosphate buffer containing 30% sucrose. The brain samples then were embedded in OCT solution, and coronal frozen sections (10 μm) were prepared. The sections were incubated with primary antibodies at 4°C overnight, including anti-β-APP (1 : 100, Abcam, Cambridge, USA), anti-ApoE (1 : 100, Abcam, Cambridge, USA), anti-NeuN (Abcam, Cambridge, USA), anti-Iba-1 (1 : 200, Wako, Osaka, Japan), and anti-P-JNK (Abcam, Cambridge, USA). Sections were then incubated with DyLight 488-conjugated goat anti-rabbit and DyLight 549-conjugated goat anti-mouse secondary antibodies (Abbkine, Redlands, USA). DAPI was used for nuclear staining. The number of positive cells was counted with Image-Pro Plus 6.0 software (Media Cybernetics, Bethesda, USA).