Glucose oxidase liquid stable reagent

For cholesterol determinations, two flies were washed in Danieau’s solution pH 7.2 (17.4 mM NaCl, 0.21 mM KCl, 0.12 mM MgSO₄+H₂0, 0.18 mM Ca(NO)₃, 1.5 mM HEPES) and homogenized in 200 μl of PBST containing 0.05% Tween 20 (Apex Bioresearch Products, Genesee Scientific El Cajon, CA #18–173). Total cholesterol was measured in triplicate using the Infinity Cholesterol Liquid Stable Reagent (Thermo Fischer Scientific Waltham, MA #T1213421). Cholesterol was quantified according to manufacturer’s instructions and were read using a SpectraMax i3 plate reader (Molecular Devices, San Jose, CA). To determine triglycerides, single flies were washed in Danieau’s solution and homogenized in 200 μL PBST. Samples were diluted 1:10 in PBST and total triglyceride was measured in triplicate using the Infinity Triglyceride reagent (Thermo Fischer Scientific Waltham, MA #TR22421). Triglyceride was quantified according to manufacturer’s instructions using a SpectraMax i3 plate reader. Glucose was determined using single flies washed in Danieau’s solution and homogenized in 500 μl of 100 mM maleate buffer, pH 5.5 (Alfa Aesar Haverhill, MA #J62049). Total glucose was measure in triplicate using the Glucose Oxidase Liquid Stable Reagent (Thermo Fischer Scientific Waltham, MA TR15221). Glucose was measured according to manufacturer’s instructions and read using a SpectraMax i3 plate reader.

Plasma levels of triglycerides, cholesterol, and glucose were measured in ZLN and ZOF rats before (control) and 3 days after mt-tat-ExoIII (ZLN) or mt-tat-EndoIII treatment (ZOF). Triglyceride Liquid Stable Reagent, Cholesterol Liquid Stable Reagent and Glucose Oxidase Liquid Stable Reagent from Infinity (Thermo Fisher Scientific, Waltham, MA, USA) were used according to the manufacturer’s instructions.