Male mice at 8 weeks of age were deprived of food for 16 hours before treatment. APAP (Sigma-Aldrich) was dissolved at a concentration of 10 mg/ml in phosphate-buffered saline that had been warmed to body temperature and was then injected intraperitoneally at a dose of 300 mg/kg. Blood was collected from the tail vein or inferior vena cava for isolation of serum by centrifugation. The activities of AST and ALT in serum were measured with Transaminase CII Test (Wako), and the serum concentrations of total cholesterol and free cholesterol were measured with Cholesterol E Test (Wako) and Free Cholesterol E Test (Wako), respectively. Glutathione concentrations in liver extracts were measured with the GSSG/GSH Quantification Kit (Dojindo).
Free cholesterol e test
Manufactured by Fujifilm
Sourced in Japan
The Free Cholesterol E Test is a laboratory equipment product designed to measure the concentration of free cholesterol in a sample. It provides a quantitative analysis of free cholesterol levels without extrapolation on intended use.
Automatically generated - may contain errors
Lab products found in correlation
Cholesterol e test, by Fujifilm (3 mentions)
Triglyceride e test, by Fujifilm (2 mentions)
Gssg gsh quantification kit, by Dojindo Laboratories (1 mentions)
Transaminase cii test, by Fujifilm (1 mentions)
Transaminase cii test wako, by Fujifilm (1 mentions)
Glucose cii test, by Fujifilm (1 mentions)
Trypan blue stain 0.4, by Thermo Fisher Scientific (1 mentions)
4 protocols using free cholesterol e test
1
APAP-induced liver injury model
2
Hepatic Lipid Extraction and Biomarker Analysis
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Hepatic lipids were extracted as described previously [10 (link)]. Serum and/or tissue levels of triglyceride, TC, free cholesterol, glucose, AST, and ALT were determined by Triglyceride E test Wako, Cholesterol E test Wako, Free cholesterol E test Wako, Glucose C II test Wako, and Transaminase C II test Wako (Wako Pure Chemical Industries, Osaka, Japan). Insulin and leptin levels were measured using a rat insulin enzyme-linked immunosorbent assay (ELISA) kit and a Leptin ELISA kit (Morinaga Institute of Biological Science, Yokohama, Japan).
3
Intracellular Lipid Quantification
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Intracellular total cholesterol, free cholesterol, and triglyceride analyses were carried out using the cholesterol E test, the free cholesterol E test, and the triglyceride E test, respectively (Wako Chemicals), according to the manufacturer’s protocol.
4
Cholesterol Accumulation in Kidney Cells
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LLC-PK1 and MES 13 were incubated with or without βVLDL. The cells were dyed with Trypan blue stain 0.4% (Life Technologies Corporation, Carlsbad, CA, USA) according to the manufacturer’s protocol and the cell number was counted to evaluate cellular proliferation.
For cholesterol accumulation, LLC-PK1 and MES 13 were treated with βVLDL for two days. After the βVLDL treatment, cells were collected and lipids were extracted using hexane: isopropanol (2:1, v/v). TC was measured using a Cholesterol E-test (Wako Pure Chemical Industries, Ltd, Osaka-shi, Japan), free cholesterol (FC) was measured using a free Cholesterol E-test (Wako Pure Chemical Industries, Ltd) and cholesterol ester (CE) was calculated by subtracting FC from TC.
For cholesterol accumulation, LLC-PK1 and MES 13 were treated with βVLDL for two days. After the βVLDL treatment, cells were collected and lipids were extracted using hexane: isopropanol (2:1, v/v). TC was measured using a Cholesterol E-test (Wako Pure Chemical Industries, Ltd, Osaka-shi, Japan), free cholesterol (FC) was measured using a free Cholesterol E-test (Wako Pure Chemical Industries, Ltd) and cholesterol ester (CE) was calculated by subtracting FC from TC.
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