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Autokit total ketone bodies

Manufactured by Fujifilm
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Sourced in United States, Japan

Autokit Total Ketone Bodies is a lab equipment product manufactured by Fujifilm. It is designed to measure the total concentration of ketone bodies in a sample. The device provides an automated, quantitative analysis of ketone bodies, which can be useful in various clinical and research applications.

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Ketone bodies (3 citations)

8 protocols using autokit total ketone bodies

1

Metabolic Profiling of Plasma and Liver

Cited 1 time
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Plasma TG, glycerol, cholesterol, NEFA, ketone bodies, ALT, insulin, and FGF21 concentrations were determined using assay kits (Serum Triglyceride Determination Kit, Sigma; Amplex Red Cholesterol Assay Kit, Life technologies; NEFA-HR (2), Wako Chemicals; Autokit Total Ketone Bodies, Wako Chemicals; ALT Kit, Bio-Quant; Ultra Sensitive Mouse Insulin ELISA Kit, Crystal Chem; Mouse/Rat FGF-21 Quantikine ELISA Kit, R&D Systems; Human FGF-21 Quantikine ELISA Kit, R&D Systems). Blood glucose levels were measured using Ascensia Breeze 2 Blood Glucose Monitoring System (Bayer). Lipids were extracted from liver tissues with chloroform/methanol mixture (2:1 v/v), as described previously66 (link).
Park J.G., Xu X., Cho S., Hur K.Y., Lee M.S., Kersten S, & Lee A.H. (2016). CREBH-FGF21 axis improves hepatic steatosis by suppressing adipose tissue lipolysis. Scientific Reports, 6, 27938.
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2

Biochemical Profiling of Fasted Mice

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Mice were fed ad libitum or fasted for 6 hr unless indicated otherwise. Blood was collected from a tail nick with a heparinized Microhematocrit capillary tube (#22-362-566; Fisher Scientific, Pittsburgh, PA) for plasma or a plain capillary (# 22-362-574; Fisher Scientific) for serum. Glucose was assayed with PGO enzymes (#P7119; Sigma) plus o-dianisidine (#F5803; Sigma). In serum samples, triglyceride was assayed with Infinity Triglycerides Liquid Stable Reagent (#TR22421; Thermo Scientific, Waltham, MA), NEFAs with HR Series NEFA-HR (2) (#999-34691; #995-34791; #991-34891; #993-35191; Wako, Richmond, VA), glycerol with Free Glycerol Reagent (#F6428; Sigma), and total ketone bodies with Autokit Total Ketone Bodies (#415-73301; #413-73601; Wako). ELISA kits were used to determine insulin (#EZRMI-13K; Millipore, Billerica, MA), and C-peptide2 (Millipore EZRMCP2-21K). For lipoprotein fractionation, fresh plasma was pooled and subjected to FPLC and assays for triglycerides and cholesterol at the UTSW Mouse Metabolic Phenotyping Core.
Ye R., Holland W.L., Gordillo R., Wang M., Wang Q.A., Shao M., Morley T.S., Gupta R.K., Stahl A, & Scherer P.E. (2014). Adiponectin is essential for lipid homeostasis and survival under insulin deficiency and promotes β-cell regeneration. eLife, 3, e03851.
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3

Plasma and Liver Metabolic Profiling

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Plasma triglycerides, non-esterified fatty acids, ketone bodies, alanine aminotransferase, and FGF21 concentrations were determined using assay kits (Serum Triglyceride Determination Kit, Sigma; NEFA-HR (2), Wako Chemicals; Autokit Total Ketone Bodies, Wako Chemicals; ALT Kit, Bio-Quant; Mouse/Rat FGF-21 Quantikine ELISA Kit, R&D Systems; Human FGF-21 Quantikine ELISA Kit, R&D Systems). Lipids were extracted from liver tissues with chloroform/methanol mixture (2:1 v/v), as described previously [45 (link)].
Ruppert P.M., Park J.G., Xu X., Hur K.Y., Lee A.H, & Kersten S. (2019). Transcriptional profiling of PPARα−/− and CREB3L3−/− livers reveals disparate regulation of hepatoproliferative and metabolic functions of PPARα. BMC Genomics, 20, 199.
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4

Plasma and Tissue Analysis in Mice

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Mice were euthanized by CO2 following the recommended procedure of the American Veterinary Medical Association (AVMA Euthanasia Panel Report). Plasma was obtained from blood samples collected with Microvette CB 300LH (SARSTEDT, Nümbrecht, Germany). Tissues were resected, snap-frozen, and stored at −80°C until further analysis. Plasma triglyceride, NEFA, ketone bodies, ALT, insulin, FGF21, albumin and haptoglobin concentrations were determined using commercial assay kits (Serum Triglyceride Determination Kit, Sigma; NEFA-HR (2 (link)), Wako Chemicals; Autokit Total Ketone Bodies, Wako Chemicals; Ultra-Sensitive Mouse Insulin ELISA Kit, Crystal Chem; Mouse/Rat FGF-21 Quantikine ELISA Kit, R&D Systems; Mouse albumin and haptoglobin ELISA kit, Abcam). Plasma Albumin and Haptoglobin levels Lipids were extracted from liver tissues with chloroform/methanol mixture (2:1 v/v) and quantitated using Serum Triglyceride Determination Kit (Sigma), as described previously (28 (link)). All assays were performed according to the manufacturer’s instructions. Blood glucose levels were measured using Ascensia Breeze 2 Blood Glucose Monitoring System (Bayer).
Xu X., Krumm C., So J.S., Bare C.J., Holman C., Gromada J., Cohen D.E, & Lee A.H. (2018). Preemptive activation of the integrated stress response protects mice from diet-induced obesity and insulin resistance via fibroblast growth factor 21 induction. Hepatology (Baltimore, Md.), 68(6), 2167-2181.
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5

Serum β-hydroxybutyrate Measurement via Enzymatic Assay

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β-hydroxybutyrate was measured for 180 serum samples. The Autokit Total Ketone Bodies (Wako Pure Chemical Industries Ltd., Richmond, VA, USA), a cyclic enzymatic method based on the oxidation of BHBA to acetoacetate by BHBA dehydrogenase, was chosen to measure serum BHBA due to its high sensitivity and high specificity.
Foditsch C., Pereira R.V., Ganda E.K., Gomez M.S., Marques E.C., Santin T, & Bicalho R.C. (2015). Oral Administration of Faecalibacterium prausnitzii Decreased the Incidence of Severe Diarrhea and Related Mortality Rate and Increased Weight Gain in Preweaned Dairy Heifers. PLoS ONE, 10(12), e0145485.
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6

Tissue Analysis of Metabolic Markers

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Tissues were dissected and fixed in 4% paraformaldehyde overnight. Paraffin processing, embedding, sectioning, and standard hematoxylin and eosin (H&E) staining were performed by the histology core at UTSW. Images were acquired using Coolscope (Nikon). Plasma glucose levels were determined by an oxidase-peroxidase assay (Sigma). Serum free fatty acid and ketone bodies levels were determined by NEFA-HR and Autokit Total Ketone Bodies (Wako Pure Chemical Industries). Plasma and tissue uridine concentrations were examined by HPLC-MS/MS at UTSW Mouse Metabolic Phenotyping Core as described [4] .
Deng Y., Wang Z.V., Gordillo R., Zhu Y., Ali A., Zhang C., Wang X., Shao M., Zhang Z., Iyengar P., Gupta R.K., Horton J.D., Hill J.A, & Scherer P.E. (2018). Adipocyte Xbp1s overexpression drives uridine production and reduces obesity. Molecular Metabolism, 11, 1-17.
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7

Colorimetric Ketone Body Quantification

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Plasma total ketone body concentration was measured using a colorimetric assay (Autokit Total Ketone Bodies; Wako Pure Chemicals) as described previously 15) .
Nishimura S., Inai M., Takagi T., Nonaka Y., Urashima S., Honda K., Aoyama T, & Terada S. (2017). Preventive Effects of the Dietary Intake of Medium-chain Triacylglycerols on Immobilization-induced Muscle Atrophy in Rats. Journal of oleo science, 66(8).
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8

Lipid and Glucose Metabolism Analysis

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Plasma and tissue levels of TG, phospholipids (PL), free cholesterol (FC), and cholesteryl esters (CE) were determined by a chemical commercial kit (Weiner Lab, Rosario, Argentina). For tissue lipid levels, lipids were extracted previous to the determination by the Folch method [11] . Blood glucose was monitored using blood glucose strips and the Accu-Check glucometer (Roche Diagnostics, Vienna, Austria). Plasma insulin concentration was measured by ELISA (Millipore Corp., MA). Plasma fatty acids (FA) levels were determined using NEFA C commercial kit according to the manufacturer's protocol (Wako, Japan). Hepatic enzymes were measured using a commercial kit (Biotron Diagnostic, CA). Ketone bodies were measured according to manufacturer's instructions (Wako, Japan). Plasma β-hydroxybutyrate levels were measured via the cyclic enzymatic method using Autokit Total Ketone bodies (Wako, Japan).
Quiroga A.D., Ceballos M.P., Parody J.P., Comanzo C.G., Lorenzetti F., Pisani G.B., Ronco M.T., Alvarez M.L, & Carrillo M.C. (2016). Hepatic carboxylesterase 3 (Ces3/Tgh) is downregulated in the early stages of liver cancer development in the rat. Biochimica et biophysica acta, 1862(11).
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