TGs (Stanbio, St. Boerne, TX), cholesterol (Wako, Mountain View, CA), β-hydroxybutyrate (Stanbio), alanine aminotransferase (ALT; Stanbio), and NEFAs (Wako) were measured using enzymatic colorimetric assays. Liver TGs were measured in Etoh:KOH lipid extracts. For liver ceramide analysis, liver was homogenized in RIPA buffer and quantitated using the Pierce™ BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA). Samples were analyzed by mass spectrometry for ceramide content at the metabolomics cores at Stony Brook University School of Medicine and at the Medical University of South Carolina. Whole and fractionated liver samples were normalized to protein and equal serum volumes were measured. Isolation of LDs, lysosomes, and ER for ceramide analysis was performed as described (26 ).
Cholesterol
Manufactured by Fujifilm
Sourced in Japan, United States
The Cholesterol lab equipment is a device used to measure the level of cholesterol in a person's blood. It provides quantitative analysis of total cholesterol, HDL cholesterol, and LDL cholesterol levels.
Automatically generated - may contain errors
Lab products found in correlation
Methanol, by Fujifilm (3 mentions)
7 dehydrocholesterol, by Merck Group (3 mentions)
Sodium taurocholate, by Fujifilm (3 mentions)
Palmitic acid, by Fujifilm (2 mentions)
Stearic acid, by Fujifilm (2 mentions)
Linoleic acid, by Fujifilm (2 mentions)
Sphingomyelin, by Fujifilm (2 mentions)
Alexa 488 conjugated secondary antibody, by Abcam (2 mentions)
Alexa fluor 568 conjugated phalloidin, by Thermo Fisher Scientific (2 mentions)
Chicken anti gfp ab13970 antibody, by Abcam (2 mentions)
Rat anti e cadherin 131900 antibody, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 647 conjugated rabbit anti cleaved caspase 3 9602s antibody, by Cell Signaling Technology (2 mentions)
Alexa fluor 568 and 647 conjugated secondary antibodies, by Thermo Fisher Scientific (2 mentions)
α linolenic acid, by Fujifilm (2 mentions)
Nile red, by Fujifilm (2 mentions)
Acetylsalicylic acid aspirin, by Merck Group (2 mentions)
Trimetazidine, by Abcam (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Acetic acid, by Fujifilm (2 mentions)
Sodium hydroxide (naoh), by Fujifilm (2 mentions)
50 protocols using cholesterol
1
Liver Lipid Metabolism Profiling
2
Plasma Cholesterol Analysis in Mice
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At the end of study, fresh EDTA plasma (50 μl) from fasted (6 h) mice was separated by fast performance liquid chromatography using an AKTA purifier and a Superose 6 column. A constant flow rate of 0.4 ml min−1 was used to collect 700 μl fractions. A 125 μl aliquot of each fraction was used to measure total cholesterol enzymatically in samples on a 96-well microtitre plate with 75 μl of two times concentrated reagents (triglyceride, RocheDiagnostics, Laval, Quebec; cholesterol, WAKO Diagnostics, Richmond, VA; and standards, Randox, Crumlin, Co. Antrim, UK).
3
C. elegans Maintenance and Synchronization
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C. elegans N2 Bristol was used in this study and maintained at 20°C on
NGM plates. The plates contained 3 g NaCl, 17 g of agar, 2.5 g peptone (Bacto, Gibco,
Thermo Fisher Scientific), 25 mL 1 mol·L−1 KPO4 buffer, 1 mL
1 mol·L−1 MgSO4, 1 mL 1 mol·L−1 CaCl2, 1 mL
5 mg·mL−1 cholesterol (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan)
in ethanol, and 975 mL H2O and were seeded with OP50. The C.
elegans strain was provided by the Caenorhabditis Genetics Center (CGC). For
synchronization of the worms, 10 adult hermaphrodites were transferred to fresh NGM plates
seeded with OP50 and cultured at 20°C for 6 hr. After the adult worms were removed, eggs
that had been deposited onto the NGM plate were grown to adulthood at 20°C [12 (link)].
NGM plates. The plates contained 3 g NaCl, 17 g of agar, 2.5 g peptone (Bacto, Gibco,
Thermo Fisher Scientific), 25 mL 1 mol·L−1 KPO4 buffer, 1 mL
1 mol·L−1 MgSO4, 1 mL 1 mol·L−1 CaCl2, 1 mL
5 mg·mL−1 cholesterol (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan)
in ethanol, and 975 mL H2O and were seeded with OP50. The C.
elegans strain was provided by the Caenorhabditis Genetics Center (CGC). For
synchronization of the worms, 10 adult hermaphrodites were transferred to fresh NGM plates
seeded with OP50 and cultured at 20°C for 6 hr. After the adult worms were removed, eggs
that had been deposited onto the NGM plate were grown to adulthood at 20°C [12 (link)].
4
Serum Metabolite Profiling in Mice
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At sacrifice, blood was collected from the heart, and serum was isolated after centrifugation for 10 min at 5,000 g. TAG (SB-2100-430; Stanbio Laboratory, Boerne, TX), glucose (997-03001; Wako Diagnostics, Lexington, MA), ketone bodies (415-73301; Wako Diagnostics, Lexington, MA), NEFAs (999-34691; Wako Diagnostics, Lexington, MA), and cholesterol (999-02601; Wako Diagnostics, Lexington, MA) were measured using 96-well plate formats as per the manufacturer's instructions.
5
Lipoprotein Cholesterol and Liver Lipid Analysis
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4 h-fasting plasma total cholesterol (TC), free cholesterol (FC) (Wako), and triglyceride (TG) (Roche) were determined by enzymatic analysis. Plasma was fractionated by FPLC to determine cholesterol distribution among the lipoprotein classes (32 (link)). Liver lipids were extracted with chloroform: methanol (2:1), and the extract was used for enzymatic assays (33 (link)) (cholesterol, Wako; TG, Roche), and lipids were normalized to wet liver weight. Macrophage cholesterol content was measured by gas-liquid chromatography (34 (link)) and normalized to cellular protein (34 (link)).
6
Rabbit diet-induced hyperlipidemia model
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Japanese white rabbits were provided by the Laboratory Animal Center of Xi'an Jiaotong University. A total of 20 male rabbits (16 weeks old, about 2.0 kg body weight) were randomly divided into three groups: The control group (n=4) was fed with a chow diet, HCD group (n=8) was fed with a chow diet supplemented with 1% Cholesterol and high fat and high Cholesterol diet (HFCD) group (n=8) was fed a chow diet containing 6.7% lard and 1% Cholesterol for 12 weeks. Rabbits were fed with a restricted diet (100 g/day/animal) with free access to water.
Cholesterol was purchased from Wako Pure Chemical Industries, Ltd., (Osaka, Japan). The experimental protocols were approved by the Animal Administration Committee of Xi'an Medical University (Shaanxi, China) and performed according to the Xi'an Medical University Guidelines for Animal Experimentation and the Guide for the Care and Use of Laboratory Animals Published by the US National Institutes of Health (NIH publication no. 85-23, revised 1996).
Cholesterol was purchased from Wako Pure Chemical Industries, Ltd., (Osaka, Japan). The experimental protocols were approved by the Animal Administration Committee of Xi'an Medical University (Shaanxi, China) and performed according to the Xi'an Medical University Guidelines for Animal Experimentation and the Guide for the Care and Use of Laboratory Animals Published by the US National Institutes of Health (NIH publication no. 85-23, revised 1996).
7
Immunolabeling of Apoptosis and Cytoskeleton
8
Fluorescent Liposome CO2 Synthesis
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Egg-derived lecithin (No. 124–05031, Fujifilm Wako Pure Chemicals), cholesterol (No. 034–03002, Fujifilm Wako Pure Chemicals), sodium hydroxide (No. 192–15985, Fujifilm Wako Pure Chemicals), 5(6)-carboxyfluorescein (No. 21877, Sigma-Aldrich), 1 M Tris-HCl (pH 7.5) (No.318–90225, Fujifilm Wako Pure Chemicals), ethanol (No.057–00456, Fujifilm Wako Pure Chemicals), and carbon dioxide (purity > 99.95 %, Fujii Bussan) were used to synthesize fluorescent liposomes that physically load CO2. Monoethanolamine (MEA, No. 016–12453, Fujifilm Wako Pure Chemicals) was used to synthesize fluorescent liposomes that chemically load CO2, while 1 M Tris-HCl (pH 7.5) was used as a buffer.
Lecithin, cholesterol, sodium hydroxide, and 5(6)-carboxyfluorescein (5,6-CF), were used to synthesize fluorescent liposomes using the Bangham method, with Tris-HCl (1 M, pH 7.5) as the buffer solution. Methanol (No. 131–01826, Fujifilm Wako Pure Chemicals) and acetic acid (No.017–00273, Fujifilm Wako Pure Chemicals) were used to prepare the mobile phase for high-performance liquid chromatography (HPLC) analysis.
Lecithin, cholesterol, sodium hydroxide, and 5(6)-carboxyfluorescein (5,6-CF), were used to synthesize fluorescent liposomes using the Bangham method, with Tris-HCl (1 M, pH 7.5) as the buffer solution. Methanol (No. 131–01826, Fujifilm Wako Pure Chemicals) and acetic acid (No.017–00273, Fujifilm Wako Pure Chemicals) were used to prepare the mobile phase for high-performance liquid chromatography (HPLC) analysis.
9
Clodronate Liposome Preparation for Macrophage Depletion
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Clodronate liposomes were prepared as below. To generate a thin layer film, 10 mg of phosphatidylcholine from egg (Avanti Polar Lipids) and 0.16 mg of cholesterol (Wako Pure Chemicals) were dissolved in 1 ml of chloroform in a glass tube, followed by evaporation of the chloroform using nitrogen gas24 (link). The tube containing the film was dried in a desiccator overnight. Clodronate disodium (Sigma) dissolved by a concentration of 50 mg ml−1 in 1 ml of PBS was added to the tube containing the film and clodronate liposomes were generated by vortexing. Control liposomes were prepared in PBS. Liposome-containing solutions were frozen and thawed three times, and subsequently passed through an extruder (Avanti Polar Lipids) with a 400 nm membrane. After centrifugation at 10 000g for 15 min, liposomes were suspended in 1 ml of sterilized PBS. To deplete macrophages, 0.1 ml clodronate liposome solution was injected into mice via the tail vein at 6 h after HTVi.
10
Serum Biomarkers Analysis in Animal Model
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