T46 antibody against total tau

Vitreous samples were centrifuged for 15 min at 12,000 rpm to separate the cellular contents, aliquoted at 100 μl, frozen at − 80 °C, and then used for Aβ, t-tau, and p-tau181 measurements. Briefly, assays were run per the manufacturer’s instructions and in duplicate for beta-amyloid 1–40 and 1–42 (Meso Scale Discovery (MSD), Rockville, MD, #K15200E-2), tau phosphorylated at threonine 181 (p-tau181), and t-tau (MSD #K15121D-2), using capturing antibody AT270 against p-tau181 (Thermo Scientific #MN1050) and T46 antibody against total tau as the detecting antibody (Thermo Scientific). Neuroinflammatory cytokines were measured using Neuroinflammation Panel 1 (K15210G, MSD). Samples were diluted 1:2 for pro-inflammatory panel 1 (IFN-γ, IL-10, IL-12p70, IL-13, IL-1β, IL-2, IL-4, IL-6, IL-8, TNF-α), cytokine panel 1 (IL-12/IL-23p40, IL-15, IL-16, IL-17A, IL-1α, IL-5, IL-7, TNF-β, VEGF-A), and angiogenesis panel 1 (basic FGF, VEGFR-1/Flt-1, Tie-2, VEGF-C, VEGF-D), or 1:5 for vascular injury panel 2 (SAA, CRP, VCAM-1, ICAM-1). Sulfo-tag-conjugated anti-mouse secondary antibody (MSD) was used for signal detection by the MSD platform, and an MSD SECTOR S 600 Imager was used to measure the analyte levels.