Autokit 3 hb

Manufactured by Fujifilm

Sourced in Japan

The Autokit 3-HB is an automated instrument designed for the quantitative determination of 3-hydroxybutyrate (3-HB) in biological samples. It utilizes enzymatic methods to measure 3-HB concentrations, providing accurate and reliable results. The Autokit 3-HB is intended for use in clinical and research settings where the assessment of 3-HB levels is required.

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Wako Autokit 3-HB (6 citations)

9 protocols using autokit 3 hb

Plasma FGF21 and 3-HB Quantification

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Trunk blood from ad libitum fed mice was collected for plasma preparation. Plasma FGF21 was assessed by FGF21-ELISA (Millipore, Billerica, MA). 3-Hydroxybutyrate (3-HB) was quantified using the colorimetric Autokit 3-HB (Wako, Richmond, VA).

Stemmer K., Zani F., Habegger K.M., Neff C., Kotzbeck P., Bauer M., Yalamanchilli S., Azad A., Lehti M., Martins P.J., Müller T.D., Pfluger P.T, & Seeley R.J. (2015). Fibroblast growth factor 21 is not required for glucose homeostasis, ketosis and tumour suppression associated to ketogenic diets in mice. Diabetologia, 58(10), 2414-2423.

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Comprehensive Metabolic Profiling Assay

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Blood glucose levels were determined using an automatic glucose monitor (One Touch, LifeScan). In addition, commercially available kits were used to measure serum non‐esterified fatty acids (NEFA; NEFA‐HR, Wako), glycerol/triglyceride (TG; TR‐0100; Sigma‐Aldrich), ketone bodies (KB; Autokit 3‐HB, Wako), cholesterol (CH200, Randox) and insulin (80‐INSMS‐E01, Alpco) essentially according to manufacturer's instructions. All samples were loaded in order to fit within the assay range of the reagents supplied. Acylcarnitines were determined in serum by electrospray ionisation tandem mass spectrometry (ESI‐MS/MS) according to a modified method as previously described (Sauer et al, 2006), using a Quattro Ultima triple quadrupole mass spectrometer (Micromass, Manchester, UK) equipped with an electrospray ion source and a Micromass MassLynx data system.

Fuhrmeister J., Zota A., Sijmonsma T.P., Seibert O., Cıngır Ş., Schmidt K., Vallon N., de Guia R.M., Niopek K., Berriel Diaz M., Maida A., Blüher M., Okun J.G., Herzig S, & Rose A.J. (2016). Fasting‐induced liver GADD45β restrains hepatic fatty acid uptake and improves metabolic health. EMBO Molecular Medicine, 8(6), 654-669.

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Plasma βHB and FFA Levels During Intervention

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Plasma βHB and free fatty acid (FFA) concentrations were respectively determined using Autokit 3-HB and NEFA-C test kits (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan). The area under the curve (AUC) values for plasma βHB and free fatty acid concentrations above the baseline during the 8-week intervention were determined using the trapezoidal rule.

Fukazawa A., Koike A., Karasawa T., Tsutsui M., Kondo S, & Terada S. (2020). Effects of a Ketogenic Diet Containing Medium-Chain Triglycerides and Endurance Training on Metabolic Enzyme Adaptations in Rat Skeletal Muscle. Nutrients, 12(5), 1269.

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Serum Metabolic Profiling Protocol

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Serum total ketone body (TKB), βOHB (Autokit 3-HB) (Wako) and glucose assays were performed according to
manufactures protocols. GTT test was performed as described previously (Cotter et al.,
2014).

Puchalska P., Martin S.E., Huang X., Lengfeld J.E., Daniel B., Graham M.J., Han X., Nagy L., Patti G.J, & Crawford P.A. (2018). Hepatocyte-macrophage acetoacetate shuttle protects against tissue fibrosis. Cell metabolism, 29(2), 383-398.e7.

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Plasma and Serum Biomarker Analysis

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The energy substrates and biomarkers in the plasma and serum were determined using kits obtained from Fujifilm Wako Pure Chemical Corporation (Osaka, Japan) ( Autokit 3-HB, NEFA C-Test Wako, Triglyceride E-Test Wako, La-bAssay Cholesterol, Glucose C2 Test Wako, Transaminase C2 Test Wako, LabAssay Creatinine, A/G B-Test Wako) .

Fukazawa A., Karasawa T., Yokota Y., Kondo S., Aoyama T, & Terada S. (2021). The Safety of Very-long-term Intake of a Ketogenic Diet Containing Medium-chain Triacylglycerols. Journal of oleo science, 70(7).

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Metabolic Biomarkers Quantification

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Triglycerides and total cholesterol were measured using Thermo Scientific Infinity Reagents (TR22421, TR13421). Insulin and leptin were quantified by ELISA (Crystal Chem #90080, #90030). β-Hydroxybutyrate was measured using Autokit 3-HB from Wako Diagnostics (Cat. No #417-73501).

Petroni G., Spring S., Schleifer K.H., Verni F, & Rosati G. (2000). Defensive extrusive ectosymbionts of Euplotidium (Ciliophora) that contain microtubule-like structures are bacteria related to Verrucomicrobia. Proceedings of the National Academy of Sciences of the United States of America, 97(4), 1813-1817.

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Comprehensive Metabolic Profiling of Mice

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For serum glucose determination, blood was collected from the tail tip and glucose was measured using Glucocard strips (A. Meranini Diagnosis). For serum isolation, blood was collected from post-mortem heart puncture. Serum insulin (Ultra Sensitive Mouse Insulin ELISA kit; Crystal Chem Inc.), Igf1 (Mouse/Rat IGF-1 ELISA; Demeditec), leptin (Crystal Chem Inc.) and adiponectin levels (Invitrogen) were measured by ELISA following the manufacturer’s instructions. Serum triglycerides (Serum Triglyceride Determination Kit; Sigma), free fatty acids (Wako NEFA C Kit; Wako Chemicals) and ketone bodies (Autokit 3-HB; Wako Chemicals) were quantified by colorimetric assay. Alanine aminotransferase (ALT) levels were measured using the ABX PENTRA 400 clinical chemical analyser (Horiba ABX Diagnostics).

Lopez-Guadamillas E., Fernandez-Marcos P.J., Pantoja C., Muñoz-Martin M., Martínez D., Gómez-López G., Campos-Olivas R., Valverde A.M, & Serrano M. (2016). p21Cip1 plays a critical role in the physiological adaptation to fasting through activation of PPARα. Scientific Reports, 6, 34542.

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Evaluating Glucose Metabolism in Mice

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Mice were fasted overnight (16 h) and orally administered glucose solution (2 g/kg body weight) for the analysis of glucose tolerance. Fed and fasting blood glucose levels were measured through the tail blood using Antsense III (HORIBA, Kyoto, Japan) between 9:00 and 11:00 a.m. The levels of HbA_1c in whole blood, glycated serum proteins in serum, and 3-hydroxybutyric acid in plasma were determined using Direct Enzymatic HbA_1c Assay (Diazyme Laboratories, Poway, CA, USA), Glycated Serum Protein Assay (Diazyme Laboratories), and Autokit 3-HB (FUJIFILM Wako, Osaka, Japan), respectively, according to the manufacturers' instructions. To determine insulin levels, insulin was extracted from the frozen pancreas using 70% (v/v) ethanol containing 0.18 N HCl. The neutralized samples were quantified using the insulin high-range HTRF kit (PerkinElmer—Cisbio, Waltham, MA, USA) according to the manufacturer's instructions.

Kitakaze K., Oyadomari M., Zhang J., Hamada Y., Takenouchi Y., Tsuboi K., Inagaki M., Tachikawa M., Fujitani Y., Okamoto Y, & Oyadomari S. (2021). ATF4-mediated transcriptional regulation protects against β-cell loss during endoplasmic reticulum stress in a mouse model. Molecular Metabolism, 54, 101338.

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Plasma metabolite and insulin analysis

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Plasma concentrations of glucose (Autokit Glucose, Wako Chemicals USA Inc., Richmond, VA), NEFA [HR Series NEFA-HR(2), Wako Chemicals USA Inc.], and BHB (Autokit 3-HB, Wako Chemicals USA Inc.) were determined by colorimetric methods, and the inter-and intraassay coefficients of variation were 6.6 and 3.2%, 6.1 and 2.3%, and 7.8 and 7.8%, respectively. Insulin concentrations in the plasma were measured by RIA (Malven et al., 1987) , and the inter-and intraassay coefficients of variation were 10 and 7.1%, respectively.

Monteiro A.P.A., Guo J.R., Weng X.S., Ahmed B.M., Hayen M.J., Dahl G.E., Bernard J.K, & Tao S. (2016). Effect of maternal heat stress during the dry period on growth and metabolism of calves. Journal of dairy science, 99(5).

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