Ab230932

Plasma and brain concentrations of apo B was measured with commercial ELISA kit (Abcam ab230932) according to manufacturer’s instruction. Mouse Aβ concentrations were determined with commercial ELISA (Wako 294–62501 and 292–64501), while human Aβ was measured with Simoa Human Neurology 3-plex A (N3PA) according to manufacturer’s instruction.

Hepatic triglyceride secretion assays were performed at the University of Michigan Mouse Metabolic Phenotyping Center as previously described (Millar et al., 2005 (link)). Blood levels of glucose were measured using a glucometer (Acucheck, Roche, Basel Switzerland) and plasma levels of cholesterol (SB-1010–225, Fisher Scientific, Hampton NH) and triglycerides (10010303, Cayman Chemical, Ann Arbor MI) were determined using colorimetric assay kits. Plasma APOB levels were determined by ELISA (ab230932, abcam, Cambridge UK).

Sera were analyzed by a colorimetric assay for total cholesterol (SB-1010–225, Fisher Scientific, Hampton NH) and by ELISAs for PCSK9 (MPC900, R&D Systems, Minneapolis MN) and APOB (ab230932, abcam, Cambridge UK). Serum lipoprotein fractionation assays were performed at the University of Cincinnati Mouse Metabolic Phenotyping Center. Sera were pooled from 5 mice for each genotype and fractionated by fast liquid protein chromatography (FPLC) into 50 fractions. Cholesterol (NC9343696, Fisher, Hampton NH) and triglyceride (TR213, Randox Laboratories, Crumlin UK) content in each fraction were determined using a microliter plate enzyme-based assay. Liver function tests were performed at the University of Michigan In-Vivo Animal Core (IVAC) with sera collected from individual mice using a Liasys analyzer (AMS Alliance).