Triglyceride tg assay kit

To measure plasma levels of adiponectin and insulin, blood samples were collected directly from the heart and transferred to EDTA-coated tubes. The plasma fraction of whole blood was obtained using a refrigerated centrifuge (2,500 × g for 30 min). Assay kits (Mouse Adiponectin ELISA Kit, KMP0041, Thermo Fisher Scientific; Ultra Sensitive Mouse Insulin ELISA Kit, 90080, Crystal Chem; Total Cholesterol Assay Kit, K603, BioVision; Free Fatty Acid (FFA) Assay Kit, K612, BioVision, Triglyceride (TG) Assay Kit, ab65336, Abcam) were used according to the manufacturer’s instructions.

These were determined using the Cholesterol Assay Kit ‐HDL and LDL/VLDL (ab65390) and Triglyceride (TG) Assay Kit (ab65336) according to the manufacturer's instructions (Abcam, Cambridge, UK). The Cholesterol Assay Kit allowed separation via precipitation (using the buffer provided in the kit) of HDL and LDL/VLDL and then determination of cholesterol levels in these particles.

Rg3, insulin, dexamethasone, 3-isobutyl-1-methylxanthine (IBMX), indomethacin, and 3,3′,5-triiodothyronine (T3) were purchased from Sigma-Aldrich (St. Louis, MO, USA). A lipid (Oil-Red O) staining kit was purchased from BioVision (Mountain View, CA, USA). 3-(4,5-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution was purchased from ThermoFisher Scientific (Waltham, MA, USA). A triglyceride (TG) assay kit was obtained from Abcam (Cambridge, UK). Antibodies against PPARγ, perilipin, endoplasmic reticulum oxidoreductin 1 alpha (ERO1L), phosphorylated eukaryotic initiation factor 2 alpha (Phospho-eIF2α), phosphorylated Akt (Phospho-Akt), total Akt, cleaved caspase-3, procaspase-3, and β-actin were purchased from Cell Signaling Technology (Danvers, MA, USA). Horseradish peroxide-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).