Extraction of liver lipids and subsequent quantification of hepatic triglycerides, total cholesterol, and phosphatidylcholine were conducted using enzymatic assays as previously described [16 ,17 ]. Liver samples were delipidated in 4 mL of 2:1 chloroform to methanol (v/v) overnight. The organic solvent was dried under a constant stream of N2 prior to adding 6 mL of 2:1 chloroform to methanol. To separate the phases, 1.2 mL of 0.05% H2SO4 was added, samples were vortexed then centrifuged at 2000 rpm for 15 min. The bottom (organic) phase was recorded, and 0.5 mL of the organic phase was added to 1 mL of 1% TritonX-100 in chloroform. The samples were dried under N2 and 0.5 mL H2O was added prior to quantification of triglycerides (Wako), total cholesterol (Pointe Scientific), and choline-containing phospholipids (Sigma), per manufacturer's instructions. All standards and blanks were prepared in a similar fashion, and data were presented normalized to initial liver weights.
Total cholesterol
Manufactured by Horiba
Sourced in Poland
The Total Cholesterol analyzer is a laboratory instrument designed to accurately measure the total cholesterol levels in biological samples. It provides quantitative analysis of total cholesterol concentration without interpretation or extrapolation of the intended use.
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Lab products found in correlation
5 protocols using total cholesterol
1
Quantification of Hepatic Lipid Profiles
2
Plasma Lipid Profile Analysis
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Mice were fasted overnight and heparin or EDTA anticoagulated plasma was collected via cardiac puncture. Total cholesterol (Pointe Scientific), total triglyceride and high-density lipoprotein and low-density lipoprotein cholesterol (Sigma) levels were measured using colorimetric assays. Blood glucose levels were measured from whole blood using the Ascensia Breeze glucose analyzer (Bayer). Pfizer lipid studies were analysed with a Clinical Analyzer (C311 Roche/Hitachi).
3
Fasted Mouse Plasma Cholesterol and Triglycerides
4
Enzymatic Metabolic Profiling in Rats
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Enzymatic colorimetric assays were utilized to measure total cholesterol (Pointe Scientific, Warszawa, Poland), free cholesterol (DiaSys Diagnostic Systems GmbH, Holzheim, Germany), triacylglycerol (Wiener Lab, Rosario, Argentina), and phospholipid concentrations (Wako Diagnostics, Mountain View, CA, USA). The cholesteryl ester mass content of the lipoprotein fractions was calculated as the difference between total cholesterol and free cholesterol multiplied by 1.67. Creatinine concentrations in serum and urine were measured using the enzymatic method (Pointe Scientific, Warsaw, Poland). The glomerular filtration rate was calculated based on the creatinine clearance. Enzyme-linked immunoassay kits were utilized to measure the levels of rat proteins, including albumin (AssayPro, ERA3201-1) and nephrin (Wuhan EIAab Science Co., Ltd., E0937r Wuhan, Hubei, China), as well as metabolites such as 8-OHdG (Cayman Chemical, kit 589320-96, Ann Arbor, MI, USA) and 8-iso-PGF2α (Wuhan Fine Biotech Co., Ltd., ER1580, Wuhan, Hubei, China). Urinary metabolites and protein excretion were measured in samples collected from rats that were individually housed in metabolic cages for 24 h. Urine volume was determined using gravimetric measurements.
5
Comprehensive Lipid and Metabolite Profiling
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Liver and cecal content lipids were extracted using Lipid Extraction Kit Chloroform Free (Abcam). Total and cholesterol ester (Millipore/Sigma-Aldrich), triglycerides (Abcam), free fatty acids (Abcam), and total bile acids (Total Bile Assay (NBT Method), GenWay Biotech Inc; San Diego, CA) were measured using the extracted liver lipids. Cecal content total bile acids were measured using the extracted cecal content lipids (Total Bile Assay (NBT Method) assay, GenWay Biotech Inc; San Diego, CA). Postprandial plasma obtained at termination of studies (see above for details) was used to measure total cholesterol (Pointe Scientific), triglycerides (Pointe Scientific), free fatty acids (Abcam), ALT (Pointe Scientific). All assays were performed according to the manufacturer’s instructions.
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