Infinity tg assay tr22421

Liver TG content was measured as previously described [31 (link)]. Plasma TG content was determined using a commercial assay from Thermo Scientific and was conducted following the manufacturer’s protocol (Infinity TG assay #TR22421 and TG standards #TG22923; Norcross, GA). For Oil-Red-O staining, frozen liver tissues were cut at 5 μm, and mounted on slides. The sections were fixed with 10% formalin for 10 minutes and then the slides were washed with deionized water for 5 minutes. Fixed tissues were then rinsed with 60% isopropanol for 5 minutes. To prepare Oil-Red-O stock, 0.5 g of Oil-Red-O (Sigma-Aldrich, O0625) was mixed with 100 mL of isopropanol. To prepare Oil-Red-O working solution, 30 mL of Oil-Red-O stock was mixed with 20 mL of distilled water and filtered using a 0.24 μm vacuum filter. Samples were submerged in the working solution for 15 minutes, briefly rinsed with 60% isopropanol, and then rinsed with deionized water for 30 seconds before imaging.

Liver TG content was measured as previously described [41 (link)]. Briefly, lipids were extracted from liver samples (~50–100 mg) using chloroform:methanol (vol:vol = 2:1). Next, sulphuric acid (0.05 %) was added to split the phase. After centrifugation (1000 × g for 5 min at room temperature), the bottom/organic phase was transferred to a new glass tube and triton X-100 (1% in chloroform) was added. Samples were dried down at 60°C under nitrogen gas and water was added into each sample (1 ml of 1% Triton X-100 in chloroform: 0.5 ml of water). Liver TGs were measured using commercial enzymatic reagents (Wako Diagnostics L-Type TG M; Richmond, VA). Plasma TG content was determined using a commercial assay from Thermo Scientific (Infinity TG assay #TR22421 and TG standards #TG22923; Norcross, GA).