Ab212073

Mice were killed by an injection of 2% sodium pentobarbitone (50 mg/kg, i.p.), and the prefrontal cortex (PFC) and hippocampus were collected, then separated, and placed in a homogenizer. Homogenates were centrifuged at 10,000 g for 10 min at 4°C. Cytokines including TNF-α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, IFN-γ, and KC/GRO were measured by the V-Plex Proinflammatory Cytokine ELISA kit (Meso Scale Discovery, QuickPlex SQ120) according to the manufacturer's instructions.
At the indicated time-points, mice were deeply anaesthetized by 2% sodium pentobarbitone (50 mg/kg, i.p.); blood was collected from the right ventricle of the heart which was centrifuged at 10,000 g for 10 min at 4°C. For serum cytokine measurement, we performed serum levels of inflammatory cytokines by using the ELISA kits for rats according to the manufacturers' instructions (TNF-α (Abcam, ab212073), IL-1β (Abcam, ab197742), IL-6 (Abcam, ab213462)). Values were expressed as pg/ml.

Following pentobarbital euthanasia, mice were perfused of blood as above. Brains were then harvested and deaggregated by pressing gently through 40 μm cell strainers in 3 ml of DMEM medium. The homogenate was then centrifuged (1400 × g, 5 min, 4 °C), and supernatants were collected. IL-1β, IL-6, TNF, and amyloid β protein levels were detected using standard ELISA after comparison to recombinant standard (cytokines: ab210895, ab213749, and ab212073, Abcam, Cambridge, MA; amyloid β^{64 (link)}: capture antibody: #800701, detection antibody: #805504 or #805404, 1:100, Biolegend, San Diego, CA).