Colorimetric assay kit

Manufactured by Fujifilm

Sourced in Japan, United States

Colorimetric assay kits are analytical tools used to quantify the concentration of specific analytes in a sample. They rely on color-based detection methods to provide quantitative measurements. These kits are designed to be easy to use and provide reliable results.

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Lab products found in correlation

Fluorometric assay kit, by Cayman Chemical (1 mentions) Mouse igf1 elisa kit, by Thermo Fisher Scientific (1 mentions) Enzychrom glycogen assay kit, by BioAssay Systems (1 mentions) Mouse insulin elisa kit, by ALPCO (1 mentions) Colorimetric assay kit, by Cayman Chemical (1 mentions) Bio plex 200 system, by Bio-Rad (1 mentions) Onetouch, by LifeScan (1 mentions) Gm9 glucose analyzer, by Analox Instruments (1 mentions) Mouse ultrasensitive insulin elisa kit, by ALPCO (1 mentions) Cobas c111 analyzer, by Roche (1 mentions) Sta 612, by Cell Biolabs (1 mentions) Dxc 800 analyzer, by Beckman Coulter (1 mentions) Milliplex assay, by Merck Group (1 mentions) Cobas c111, by Roche (1 mentions)

15 protocols using colorimetric assay kit

Quantifying Hepatic Lipid Metabolism

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Plasma insulin and IGF-1 levels were measured with a Mouse Insulin ELISA Kit (catalogue no. 80-INSMSE01; ALPCO, USA) and Mouse IGF1 ELISA Kit (catalogue no. EMIGF1; Thermo Fisher Scientific), respectively. Total lipids from mouse livers or mouse primary hepatocytes were extracted using the Folch method as described previously [7 (link)]. Hepatic TG, cellular TG, plasma TG and NEFA levels were measured with colorimetric assay kits (TG: catalogue no. 461-09092; NEFA: catalogue no. 438-91691; Wako, Japan). Hepatic glycogen was measured using an EnzyChrom Glycogen Assay Kit (catalogue no. E2GN-100; BioAssay Systems, USA). Hepatic β-hydroxybutyrate (β-OH) was measured with a fluorometric assay kit (catalogue no. 700740; Cayman Chemical, USA).

Lee D.S., An T.H., Kim H., Jung E., Kim G., Oh S.Y., Kim J.S., Chun H.J., Jung J., Lee E.W., Han B.S., Han D.H., Lee Y.H., Han T.S., Hur K., Lee C.H., Kim D.S., Kim W.K., Park J.W., Koo S.H., Seong J.K., Lee S.C., Kim H., Bae K.H, & Oh K.J. (2023). Tcf7l2 in hepatocytes regulates de novo lipogenesis in diet-induced non-alcoholic fatty liver disease in mice. Diabetologia, 66(5), 931-954.

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Serum Biomarkers in Fasted Mice

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Serum levels of insulin, glucagon, gastric inhibitory polypeptide (GIP), leptin, resistin, and GLP-1 were measured in plasma isolated from mice that were fasted overnight using a multiplex bead-based assay performed on a Bio-Plex 200 system (Bio-Rad, Mississauga, ON, Canada) (Bio-Rad Laboratories, Missisissauga, Ontario, Canada) according to the manufacturer's instructions. Serum TAG, NEFA, and total cholesterol were measured using colorimetric assay kits (Wako Diagnostics, Richmond, VA, USA). Serum creatinine was measured using a colorimetric assay kit (Cayman Chemicals, Ann Arbour, MI, USA).

Fernandes M.F., Aristizabal-Henao J.J., Marvyn P.M., M'Hiri I., Wiens M.A., Hoang M., Sebastian M., Nachbar R., St-Pierre P., Diaguarachchige De Silva K., Wood G.A., Joseph J.W., Doucette C.A., Marette A., Stark K.D, & Duncan R.E. (2024). Renal tubule-specific Atgl deletion links kidney lipid metabolism to glucagon-like peptide 1 and insulin secretion independent of renal inflammation or lipotoxicity. Molecular Metabolism, 81, 101887.

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Metabolic Biomarkers Measurement Protocol

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Blood glucose levels under basal conditions and during glucose tolerance tests were measured using an automatic glucose monitor (One Touch; LifeScan, Milpitas, CA, USA). Plasma and hepatic TG, cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and non-esterified fatty acid (NEFA) levels were measured using colorimetric assay kits from Wako (Wako Pure Chemical Industries, Ltd., Osaka, Japan).

Kim G.T., Kim S.J., Park S.H., Lee D, & Park T.S. (2020). Hepatic Expression of the Serine Palmitoyltransferase Subunit Sptlc2 Reduces Lipid Droplets in the Liver by Activating VLDL Secretion. Journal of Lipid and Atherosclerosis, 9(2), 291-303.

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Metabolic Biomarker Quantification Protocol

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Blood glucose during hyperinsulinemic-clamp and glucose tolerance test was measured using the GM9 Glucose Analyzer (Analox Instruments). Lipids in tissue were extracted using the method of Bligh and Dyer55 (link) and hepatic triglyceride, cholesterol and plasma NEFA were measured by colorimetric assay kits from Wako (Wako Pure Chemical Industries, 279-75401). Plasma triglyceride, total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), ALT, and AST were measured using a Cobas c111 analyzer (Roche). Plasma insulin was measured using Mouse Ultrasensitive Insulin ELISA kits (ALPCO, 80-INSMSU-E01).

Hur J.H., Park S.Y., Dall’Armi C., Lee J.S., Di Paolo G., Lee H.Y., Yoon M.S., Min D.S, & Choi C.S. (2016). Phospholipase D1 deficiency in mice causes nonalcoholic fatty liver disease via an autophagy defect. Scientific Reports, 6, 39170.

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Cholesterol and Choline Phospholipid Quantification

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HEK293 cells were transfected with the expression vectors and incubated for 48 h. Then, lipids were extracted from the cells with chloroform/methanol (2:1). The lipid-containing solution was dried and resuspended in 2-propanol, and the levels of cholesterol and choline phospholipids were determined by colorimetric assay kits purchased from FUJIFILM Wako Pure Chemical Corporation (435-35801 for free cholesterol and 433-36201 for choline phospholipids).

Nakato M., Shiranaga N., Tomioka M., Watanabe H., Kurisu J., Kengaku M., Komura N., Ando H., Kimura Y., Kioka N, & Ueda K. (2020). ABCA13 dysfunction associated with psychiatric disorders causes impaired cholesterol trafficking. The Journal of Biological Chemistry, 296, 100166.

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Aortic Calcium and Phosphate Quantification

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To determine levels of Ca and Pi in aortas, the aortas were dried at 55 °C and then weighed. Ca and Pi were extracted with 150 mM HCl overnight at room temperature. The Ca and Pi content in the solution was measured using Colorimetric Assay kits (Wako, Osaka, Japan) according to the manufacturer’s instructions. The Ca and Pi content was determined in relation to the dry tissue weight.

Wei W., Guo X., Gu L., Jia J., Yang M., Yuan W, & Rong S. (2021). Bone marrow mesenchymal stem cell exosomes suppress phosphate-induced aortic calcification via SIRT6–HMGB1 deacetylation. Stem Cell Research & Therapy, 12, 235.

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Quantification of Hepatic Lipid Accumulation

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Hepatic fat accumulation was determined by Oil Red O staining. Briefly, liver cryosections (5 μm thick) were fixed with 4% PFA in PBS for 20 min and incubated in freshly prepared Oil Red O working solution (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan) for 10 min and counterstained with hematoxylin for 15 sec. The random images of each Oil Red O stained liver frozen tissue sections (n = 3 per group) were captured at ×200 magnification. Relative areas of steatosis, expressed as % of fat accumulation, were quantified by histomorphometry using WinROOF image analyzer (Version 7, Mitani Corp, Tokyo, Japan). Moreover, total hepatic lipid was extracted from fresh frozen liver tissues using a lipid extraction kit (STA-612; Cell biolabs, San Diego, CA, USA). Hepatic triglycerides and cholesterol were then quantitated using colorimetric assay kits, according to the manufacturer’s instructions (Fujifilm Wako).

Srisowanna N., Choijookhuu N., Yano K., Batmunkh B., Ikenoue M., Nhat Huynh Mai N., Yamaguchi Y, & Hishikawa Y. (2019). The Effect of Estrogen on Hepatic Fat Accumulation during Early Phase of Liver Regeneration after Partial Hepatectomy in Rats. Acta Histochemica et Cytochemica, 52(4), 67-75.

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Comprehensive Metabolic Profiling in Mice

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Blood fasting glucose was checked from tail-vein blood using an automatic glucose monitor (One Touch). Plasma triglycerides, and total cholesterol and liver triglyceride were measured with colorimetric assay kits (Wako, Osaka, Japan). Total liver lipids were extracted with chloroform-methanol (2:1, v/v) mixture according to the Folch method [12 (link)]. Plasma insulin levels were measured with a commercial insulin assay ELISA kit (ALPCO Diagnostics, Salem, NH, USA). Serum levels of alanine aminotransferase (ALT) were determined using a Beckman DXC 800 analyzer (Brea, CA, USA).

Cho J., Lee I., Kim D., Koh Y., Kong J., Lee S, & Kang H. (2014). Effect of aerobic exercise training on non-alcoholic fatty liver disease induced by a high fat diet in C57BL/6 mice. Journal of Exercise Nutrition & Biochemistry, 18(4), 339-346.

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Quantifying Triacylglycerol Metabolism in Hearts

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Triacylglycerol (TAG) content was assessed in the set of hearts perfused with [9,10-³H]palmitate in the perfusate (n = 6 to 7/group) as described previously (16) (link) by colorimetric assay kit (Wako Pure Chemical Industries, Richmond, Virginia), while another potion of the lipid extraction was counted with scintillation fluid to measure the incorporation of [9,10-³H]palmitate into TAG based on the specific activity of [9,10-³H]palmitate from the perfusate.

Wang W., Zhang L., Battiprolu P.K., Fukushima A., Nguyen K., Milner K., Gupta A., Altamimi T., Byrne N., Mori J., Alrob O.A., Wagg C., Fillmore N., Wang S.H., Liu D.M., Fu A., Lu J.Y., Chaves M., Motani A., Ussher J.R., Reagan J.D., Dyck J.R, & Lopaschuk G.D. (2019). Malonyl CoA Decarboxylase Inhibition Improves Cardiac Function Post-Myocardial Infarction. JACC: Basic to Translational Science, 4(3), 385-400.

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Intracellular Calcium Content Quantification

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To measure intracellular Ca content, cells were first decalcified in 0.6 mM HCl at 4 °C for 24 h. Ca released by the cells was measured with a Colorimetric Assay kit (Wako, Osaka, Japan) according to the manufacturer’s instructions. The Ca content was normalized to cell protein content using a BCA assay (Pierce, Waltham, MA, USA).

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