Rabbit anti neun monoclonal antibody

All drugs were purchased from Sigma-Aldrich. Mouse Anti-BrdU monoclonal antibody was purchased from Sigma-Aldrich. Rabbit anti-NeuN monoclonal antibody was purchased from Millipore. Rabbit anti-GFAP monoclonal was purchased from Abcam. Complete protease inhibitor cocktail tablets and phosphatase inhibitor cocktail tablets were purchased from Roche Applied Science.

Mice were anesthetized with 125mg/kg Ketamine and 10mg/kg Xylazine cocktail intraperitoneal injection. The brain was fixed by the transcardial perfusion of 4% PFA/PBS, followed by overnight incubation in an additional 4% PFA/PBS. After removing the fixative, the brain was cryoprotected with a 10–30% sucrose solution. The section was made with 30μm slices. Blocking was achieved with 4% BSA in PBS and 0.3% Triton X-100. Primary antibodies and working dilution are the following: Mouse anti-β-Amyloid antibody (1:500, clone: 6E10, Biolegend), Rabbit anti-NeuN monoclonal antibody (clone: A60, Millipore), and Rabbit anti-Iba1 antibody (Wako). The tissue was then incubated with the following fluorescence-labeled secondary antibodies; Alexa Fluor 488 Goat anti-mouse IgG (H+L) (Invitrogen), Alexa Fluor 633 Goat anti-mouse IgG (H+L) (Invitrogen), Alexa Fluor 633 Goat anti-rabbit IgG (H+L) (Invitrogen). The core-dense amyloid plaques were stained with 0.5% Thioflavin S (Sigma, T1892). The nuclei were stained with Hoechst33342 dye. The images were taken on a Zeiss AiryScan 880 confocal microscope and further analyzed with ImageJ/Fiji^{50 (link)}. Five fields per section were randomly chosen in the cortical layer V and analyzed for quantification. The average value of 5 fields was presented for each mouse.

Lithium chloride (LiCl) and bromodeoxyuridine (BrdU) were purchased from Sigma-Aldrich, and dissolved in 0.9% sterile saline at required concentrations.
Mouse Anti-BrdU monoclonal antibody was obtained from Sigma-Aldrich. Rabbit anti-NeuN monoclonal antibody was obtained from Millipore. Rabbit anti-GFAP monoclonal and rabbit anti-doublecortin (DCX) polyclonal antibodies were obtained from Abcam. Complete protease inhibitor cocktail tablets and phosphatase inhibitor cocktail tablets were purchased from Roche Applied Science.