Total HDL was isolated by sequential ultracentrifugation (density range 1.063–1.210 g/ml), using potassium bromide (KBr) gradients. HDL composition was determined by measuring the content of total cholesterol (Roche Diagnostics), TG (Roche Diagnostics), free cholesterol (Wako Chemicals, Richmond, VA, USA) phospholipid (Wako Chemicals), apoA-I (Roche Diagnostics) apoA-II (Kamiya Biomedical Company, Seattle, WA, USA), apoE (Kamiya Biomedical Company) and apoC-III (Kamiya Biomedical Company). Composition data are expressed as a % of HDL total mass. PON-1 activity in serum was measured using phenylacetate as a substrate, as described [19 (link)]. Lp-PLA2 activity was measured using 2-thio-PAF (Cayman Chemical Company, Ann Arbor, MI, USA) as a substrate, according to the manufacturer’s instructions [20 (link)]. To determine the distribution of Lp-PLA2 among the lipoprotein fractions, apoB-containing lipoproteins were precipitated from serum using dextran sulfate, as described [21 (link)].
Phospholipid
Manufactured by Fujifilm
Sourced in United States, Argentina, Germany, Spain
Phospholipid is a type of lipid molecule that is an essential component of cell membranes. It consists of a glycerol backbone, two fatty acid chains, and a phosphate group. Phospholipids play a crucial role in maintaining the structural integrity and permeability of cell membranes, allowing for the proper transport of substances in and out of cells.
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Lab products found in correlation
Total cholesterol, by Roche (1 mentions)
Free cholesterol, by Fujifilm (1 mentions)
2 thio paf, by Cayman Chemical (1 mentions)
Apoa 1, by Roche (1 mentions)
Pierce coomassie bradford protein assay kit, by Thermo Fisher Scientific (1 mentions)
Total cholesterol, by Horiba (1 mentions)
Free cholesterol, by DiaSys (1 mentions)
Triglyceride, by Merck Group (1 mentions)
Total cholesterol, by Randox (1 mentions)
Triglyceride, by Roche (1 mentions)
Triglyceride, by Cayman Chemical (1 mentions)
Rat glucagon elisa kit, by Cusabio (1 mentions)
Rat insulin elisa kit, by Merck Group (1 mentions)
8 protocols using phospholipid
1
HDL Composition and Enzyme Activities
2
Quantifying Lipoprotein Components
3
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.
4
Hepatic Lipid Extraction and Analysis
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Hepatic lipid extraction was conducted as described before (Folch et al, 1957 (link)). Total cholesterol (Randox Laboratories Ltd.), non-esterified fatty acid (FUJIFILM Wako Chemicals), phospholipid (FUJIFILM Wako Chemicals), and triglyceride (Sigma-Aldrich) levels were determined using commercially available kits according to the manufacturer’s instructions.
5
Lipoprotein Cholesterol Determination
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In plasma samples, total cholesterol, triglycerides (Roche Molecular Biochemicals, Mannheim, Germany) and phospholipids (Wako, Neuss, Germany) were determined enzymatically using commercially available kits (n=5 per group). Not all animals yielded sufficient amounts of blood for parallel assessment of IL-6 and lipoprotein profiles. Animals with sufficient plasma sample volume were selected. The cholesterol content within different lipoprotein subclasses from individual plasma samples was determined following sequential tabletop ultracentrifugation as published (Tietge et al., 2000 (link)). Cholesterol concentrations within each fraction were measured as detailed above.
6
Enzymatic Analysis of Serum and Liver Lipids
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Serum concentrations of cholesterol (Wako Diagnostics), phospholipids (Wako), triglycerides (Cayman Chemical), total bile acids (Crystal Chem), and non-esterified fatty acids (NEFA) (Wako) were determined enzymatically according to manufacturers’ instructions. These enzymatic assays were also used for the analysis of hepatic contents of cholesterol, phospholipids, and triglycerides after the extraction of lipids from liver samples, as described.16 (link) Serum activities of alanine transaminase (ALT) and aspartate transaminase (AST) were determined using the assay kits from Pointe Scientific according to the company’s instructions.
7
Serum and Liver Lipid Profiling in Mice
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After a 4 h fast, mice were anaesthetized by intraperitoneal injection with sodium pentobarbital. Blood was collected by heart puncture and liver was removed, weighed, and snap-frozen in liquid nitrogen. The collected blood was centrifuged at 3500 rpm for 10 min at 4 °C, and the serum was analyzed for total and free cholesterol, triglycerides and phospholipids concentrations using the Cholesterol/HP(Roche), the Free Cholesterol C(Wako), Triglycerides/GB kit(Roche) and phospholipids(Wako) enzymatic assay kits, respectively.
For analysis of the liver lipid composition, the lipid extract was separated from liver as described39 (link). Lipids were then quantified using the same enzymatic kits described above.
For analysis of the liver lipid composition, the lipid extract was separated from liver as described39 (link). Lipids were then quantified using the same enzymatic kits described above.
8
Metabolic Profiling of Rodent Serum
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Glucose, total cholesterol, and triglycerides (QCA, Barcelona, Spain), phospholipids (Spinreact, Girona, Spain), and non-esterified free fatty acids (NEFAs) (WAKO, Neuss, Germany) were analyzed by enzymatic colorimetric assays. Serum insulin and glucagon levels were analyzed using a rat insulin ELISA kit (Millipore, Barcelona, Spain) and a rat glucagon ELISA kit (Cusabio Biotech, Wuhan, China), respectively.
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