Lipid panel plus diskettes

Plasma levels for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and circulating levels of high-density lipoprotein (HDL), triglycerides, and cholesterol were obtained using an Abaxis Piccolo Xpress analyzer (Abbott; Abbott Park, IL, USA) with Lipid Panel Plus diskettes (catalog:400-0030; Abaxis Inc.; Union City, CA, USA). Circulating levels of plasma adipokines and cytokines were measured using a MILLIPLEX^® Mouse Adipokine Magnetic Bead Panel (Millipore Sigma, Burlington, MA, USA) for analytes interleukin 6 (IL-6), leptin, total plasminogen activator inhibitor 1 (PAI-1), tumor necrosis factor alpha (TNFα), and resistin. Data was obtained from a Luminex 100/200 System equipped with xPONENT Software (Luminex Corporate, Austin, TX, USA). Hepatic lipids were extracted using a modified Bligh and Dyer method with a 2:1 ratio of chloroform to methanol mixture (27 (link), 28 (link)). Extracted lipids were measured using Infinity Assay Reagents: Infinity Triglycerides reagent (TR22421, ThermoFisher Scientific, Middletown, VA, USA) and Infinity Cholesterol reagent (TR13421, ThermoFisher Scientific, Middletown, VA, USA). Sample absorbance at 500 nm was calculated from a standard curve constructed from clinical standards (catalog: T7531-STD, C7509-STD; Point Scientific; Canton, MI) for each analyte ran simultaneously with unknown samples.

Liver enzymes, triglycerides, glucose, cholesterol, and lipoproteins (HDL, LDL, VLDL) were measured with Lipid Panel Plus diskettes (catalog:400-0030; Abaxis Inc.; Union City, CA) on the Piccolo Xpress Chemistry Analyzer (Abbott; Abbott Park, IL). Plasma adipokines were measured with the Milliplex Map Mouse Adipokine Magnetic Bead Panels (catalog: MADKMAG-71K; EMD Millipore) on a Luminex^® 100 system (Luminex Corp.; Austin, TX). Adipokines measured included Insulin, PAI-1, Resistin, and Leptin. Analysis for each assessment was performed according to the manufacturer’s protocol. Plasma albumin levels were measured via ELISA from MyBioSource Inc. and performed according to the manufacturer’s standard protocol (catalog: MBS564063; San Diego, CA).