Triglyceride e test wako kit

Manufactured by Fujifilm

Sourced in Japan

The Triglyceride E-test Wako kit is a laboratory equipment product designed for the quantitative determination of triglycerides in biological samples. It provides a reliable and efficient method for measuring triglyceride levels.

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25 protocols using triglyceride e test wako kit

Triglyceride Quantification in Adipocytes

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The cellular
concentration of triglyceride (TG) was determined using the method
of Senarath et al.(45 (link)) with
a few modifications. Briefly, mature adipocytes obtained on the eighth
day were rinsed twice with PBS; the cells were scraped into PIPES
(Piperazine-N,N′-bis(2-ethanesulfonic acid)) buffer with 0.1% Triton X and
lysed by sonication on ice. The TG level in the lysate was measured
using the triglyceride E-test Wako kit (FUJIFILM Wako Pure Chemical
Corporation) according to the manufacture’s protocol.

Takahashi J., Takahashi N., Tadaishi M., Shimizu M, & Kobayashi-Hattori K. (2022). Valerenic Acid Promotes Adipocyte Differentiation, Adiponectin Production, and Glucose Uptake via Its PPARγ Ligand Activity. ACS Omega, 7(51), 48113-48120.

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Fenofibrate and Phenobarbital Effects on Liver

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Male C57BL/6N mice (8–12 weeks old) were maintained under a 12 h light/dark cycle and fed a conventional laboratory diet and water. Mice were fed a purified diet (CE-2; Clea Japan, Tokyo, Japan) supplemented with or without fenofibrate (0.03 or 0.1% wt/wt) and phenobarbital (0.03 or 0.1% wt/wt) for a week. Mice were intraperitoneally treated with TCPBOP (3 mg/kg dissolved in corn oil). Six hours after treatment, the mice fasted for 18 h. Livers and plasma were collected and subjected to experiments. Plasma triglyceride levels were determined using a Triglyceride E Test WAKO kit (Fujifilm Wako Pure Chemicals). All animal experiments were approved by the committees for animal experiments at Tohoku University and the University of Shizuoka and conducted in accordance with the guidelines for animal experiments at Tohoku University and the University of Shizuoka.

Shizu R., Otsuka Y., Ishii C., Ezaki K, & Yoshinari K. (2023). PPARα Induces the Expression of CAR That Works as a Negative Regulator of PPARα Functions in Mouse Livers. International Journal of Molecular Sciences, 24(4), 3953.

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Liver Lipid Extraction and Triglyceride Quantification

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Lipid extraction from the liver tissues was performed as previously described [29 (link)]. The triglyceride content in the lipid extracts was measured using the Triglyceride E-test Wako kit (WAKO) according to the manufacturer’s instructions.

Tsuzuki T., Shinozaki S., Nakamoto H., Kaneki M., Goto S., Shimokado K., Kobayashi H, & Naito H. (2015). Voluntary Exercise Can Ameliorate Insulin Resistance by Reducing iNOS-Mediated S-Nitrosylation of Akt in the Liver in Obese Rats. PLoS ONE, 10(7), e0132029.

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Hepatic Biomarker Assessment

Cited 1 time

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Plasma alanine aminotransferase (ALT) levels were measured using the Transaminase CII-Test-Wako kit (Fujifilm Wako Pure Chemical Corp.). Hepatic triglyceride contents were measured using the triglyceride E-Test-Wako kit (Fujifilm Wako Pure Chemical Corp.)^{47 (link)}. Hepatic MDA levels were measured using the Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) (ab118970, Abcam, Cambridge, UK).

Inaba Y., Hashiuchi E., Watanabe H., Kimura K., Oshima Y., Tsuchiya K., Murai S., Takahashi C., Matsumoto M., Kitajima S., Yamamoto Y., Honda M., Asahara S.I., Ravnskjaer K., Horike S.I., Kaneko S., Kasuga M., Nakano H., Harada K, & Inoue H. (2023). The transcription factor ATF3 switches cell death from apoptosis to necroptosis in hepatic steatosis in male mice. Nature Communications, 14, 167.

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Hepatic Triglyceride Quantification

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To examine triglyceride content in the liver, hepatic triglyceride assay was performed with Triglyceride E-test Wako kit (Wako Pure Chemical Industries, Tokyo, Japan) according to the manufacturer’s instruction.

Fujiwara S., Izawa T., Mori M., Atarashi M., Yamate J, & Kuwamura M. (2022). Dietary iron overload enhances Western diet induced hepatic inflammation and alters lipid metabolism in rats sharing similarity with human DIOS. Scientific Reports, 12, 21414.

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Hepatic Triglyceride Content Quantification

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After liver tissue weight was measured, frozen liver tissue was homogenized in 2 mL ice-cold isopropanol. The samples were incubated for 10 min with shaking at room temperature, and the samples were centrifuged at 3000 rpm for 10 min. 1 mL of supernatant was used to measure the hepatic triglyceride content by commercial Triglyceride E-test WAKO kit (Wako Pure Chemical Industries, Osaka, Japan). Results were normalized to the weight of each liver samples.

Takata N., Ishii K.A., Takayama H., Nagashimada M., Kamoshita K., Tanaka T., Kikuchi A., Takeshita Y., Matsumoto Y., Ota T., Yamamoto Y., Yamagoe S., Seki A., Sakai Y., Kaneko S, & Takamura T. (2021). LECT2 as a hepatokine links liver steatosis to inflammation via activating tissue macrophages in NASH. Scientific Reports, 11, 555.

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Adipogenic Differentiation of 3T3-L1 Cells

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Mouse 3T3-L1 preadipocytes were obtained from the European Collection of Cell Cultures (Wiltshire, UK). The Transcriptor First Strand cDNA Synthesis Kit and LightCycler FirstStart DNA Master^plus SYBR Green reagent were obtained from Roche Diagnostics (Indianapolis, IN, USA). TRIzol reagent and the primers for β-actin, PPARγ, and C/EBPα were purchased from Invitrogen (Carlsbad, CA, USA). Rabbit polyclonal anti-human β-actin, PPARγ, and C/EBPα antibodies and goat HRP-linked anti-rabbit IgG antibody were obtained from Cell Signaling Technology, Beverly, MA, U.S.A. Sulforaphane, allyl isothiocyanate, and a protease inhibitor cocktail were obtained from Sigma Chemical Co. (St. Louis, MO, USA). The Triglyceride E-test Wako kit was obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). The Lipid Droplets Fluorescence Assay Kit was obtained from Cayman Chemical Co. (Ann Arbor, MI, USA). All other reagents used were of analytical grade.

Sakuma S., Yasuda K., Kitahara R., Tsujimoto K., Yamashita K., Hoshino N., Fujimoto Y, & Okuhira K. (2022). Comparative Effects of Sulforaphane and Allyl Isothiocyanate on 3T3-L1 Adipogenesis. Journal of Nutrition and Metabolism, 2022, 8705163.

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Intracellular Triglyceride Quantification

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C2C12 myotubes were washed with PBS, and lipids were extracted by hexane in 2-propanol (3:2, v/v). The levels of intracellular TG were determined using the Triglyceride E-test Wako Kit (FUJIFILM Wako Pure Chemical) and normalized to the levels of total cellular protein determined using a BCA protein assay kit (Pierce), according to each manufacturer’s instructions.

Sasaki T., Watanabe Y., Kuboyama A., Oikawa A., Shimizu M., Yamauchi Y, & Sato R. (2020). Muscle-specific TGR5 overexpression improves glucose clearance in glucose-intolerant mice. The Journal of Biological Chemistry, 296, 100131.

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Serum Lipid and Liver Enzyme Profiling

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The concentrations of cholesterol, bile acids, phospholipids and TG, and the activity of alanine aminotransferase (ALT) in each sample were measured using commercially available kits according to manufacturer’s instructions. The Cholesterol E-test Wako Kit, the TBA test Wako Kit (for the determination of total bile acids), the Phospholipids C-test Wako Kit, the Triglyceride E-test Wako Kit, and the Transaminase CII-test Wako Kit (Wako Pure Chemical Industries, Tokyo, Japan) were used in this study.

Toyoda Y., Takada T., Yamanashi Y, & Suzuki H. (2019). Pathophysiological importance of bile cholesterol reabsorption: hepatic NPC1L1-exacerbated steatosis and decreasing VLDL-TG secretion in mice fed a high-fat diet. Lipids in Health and Disease, 18, 234.

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Triglyceride Quantification in Cells

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TAG measurement analysis was performed as previously described [27 (link)]. Briefly, after washing the cells with PBS, all lipids were extracted by the Folch method. Extracted lipids were resuspended in isopropanol. TAG levels were measured using a triglyceride E test Wako kit (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) according to the manufacturer’s protocol. The intracellular TAG level was normalized by the total protein level.

Kuramoto K., Yamamoto M., Suzuki S., Togashi K., Sanomachi T., Kitanaka C, & Okada M. (2021). Inhibition of the Lipid Droplet–Peroxisome Proliferator-Activated Receptor α Axis Suppresses Cancer Stem Cell Properties. Genes, 12(1), 99.

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