Triglyceride Quantification in Flies

Triglyceride assay was performed as previously described [9 (link)]. Briefly, adult flies Flies (12 per genotype, 1:1 ratio of males:females) were weighed and homogenized in PBS containing 0.1% Triton-X100 in an amount (μl) that is 8 X the total weight of flies (μg). The homogenates were centrifuged and the supernatants were removed and incubated with Infinity Triglyceride Reagent (Thermo Electron) for 30 min at 37°C. The absorbance at 540 nm was then measured and TG content was calculated from a standard curve constructed with solutions of known TG concentrations (Thermo Electron). The results were normalized to the protein concentration (μg/μl) of each sample with the Bradford assay (5000002, Bio-Rad).