Autokit glucose

Manufactured by Fujifilm

Sourced in United States, Japan, Canada

The Autokit Glucose is a laboratory equipment used for the automated quantitative determination of glucose levels in biological samples. It utilizes a colorimetric method to measure the glucose concentration accurately and efficiently.

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Spelling variants of this product

Autokit Glucose reagent (4 citations) Wako Autokit Glucose (3 citations) Autokit Glucose C2 (3 citations) Autokit glucose assay (2 citations)

41 protocols using autokit glucose

Quantification of Metabolic Biomarkers

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Serum glucose was measured using Autokit Glucose (Wako, Richmond, VA). Serum leptin was assayed using the rat leptin radioimmunoassay kit from Linco Research, Inc. (St Louis, MO) according to the manufacturer’s instructions. Serum IGF1 was measured with a radioimmunoassay using a polyclonal antibody to IGF1 after separation of IGF-binding proteins by acid ethanol extraction. Serum osteocalcin was measured using rat Gla-osteocalcin high sensitive EIA kit (Clontech, Mountain View, CA). Serum CTx was measured using rat CTx-I ELISA kit (Novateinbio, Cambridge, MA). Serum adiponectin was measured using rat total adiponectin Quantikine ELISA kit (R&D Systems, Minneapolis, MN). Serum growth hormone was measured using a rat/mouse growth hormone ELISA kit (EMB Millipore, Billerica, MA).

Turner R.T., Dube M., Branscum A.J., Wong C.P., Olson D.A., Zhong X., Kweh M.F., Larkin I.V., Wronski T.J., Rosen C.J., Kalra S.P, & Iwaniec U.T. (2015). Hypothalamic Leptin Gene Therapy Reduces Body Weight without Accelerating Age-Related Bone Loss. The Journal of endocrinology, 227(3), 129-141.

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Comprehensive Lipid Metabolism Profiling

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Infinity Cholesterol (catalog #TR13421), Infinity Triglyceride (catalog #TR22421), and TRIzol^TM (catalog #15596018) reagents were purchased from Thermo Fisher Scientific (Middletown, VA, USA). Autokit Glucose (catalog #997-03001), Phospholipids C (catalog #997-01801), and HR Series NEFA-HR(2) (catalog #999-34691, 995-34791, 991-34891, and 993-35191) kits were purchased from Fujifilm Wako Chemicals USA (Richmond, VA, USA). Omniscript RT (catalog #205113) kit was purchased from Qiagen (Germantown, MD, USA) and qPCR^TM core kit for SYBR Green I (catalog #10-SN10-05) was from Eurogentec (San Diego, CA, USA). ³H-cholesterol (catalog #NET139001MC), ¹⁴C-oleic acid (catalog #NEC317250UC), and ¹⁴C-triolein (catalog #NEC674250UC) were from PerkinElmer (Shelton, CT, USA). Poloxamer 407 (catalog #P1166) was purchased from Spectrum Chemical (New Brunswick, NJ, USA). Primary and secondary antibodies were purchased from either Cell Signaling (Danvers, MA, USA) or Abcam (Cambridge, MA, USA). All other chemicals and solvents were obtained from Fisher Scientific through its local distributor in the Kingdom of Saudi Arabia.

Otaibi A.A., Mubarak S.A., Qarni A.A., Hawwari A., Bakillah A, & Iqbal J. (2022). ATP-Binding Cassette Protein ABCC10 Deficiency Prevents Diet-Induced Obesity but Not Atherosclerosis in Mice. International Journal of Molecular Sciences, 23(22), 13813.

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Comprehensive Kidney Function Evaluation

Cited 2 times

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Haematocrit was determined using glass microcapillaries at the first time point of inulin clearance. Urinary albumin concentration was measured by ELISA (Albuwell and Creatinine Companion, Exocell, Philadelphia, PA, USA)[13 (link)–15 (link), 18 (link), 22 (link)], and 24-h urinary albumin excretion was calculated with the urine volume. Serum and urine glucose concentrations were determined using a colorimetric-enzymatic method (Autokit Glucose, Wako Diagnostics, Richmond, VA, USA). Fractional reabsorption of glucose (FR-glucose) was calculated using the serum and urine glucose concentration, urine volume, and GFR of each mouse [23 (link)]. Urinary Ang II levels were measured by a commercial ELISA kit (Bachem America, Torrence, CA, USA) according to the protocol III as described previously [15 (link), 18 (link)]. Urinary adenosine was assayed by fluorometric method (Abcam (Cambridge, MA, USA), catalogue no. ab211094). The urine was pre-treated with catalase beads (Abcam, catalogue no. 218718) and the fluorescence signal was detected by excitation 535nm and emission 587nm.

Miyata K.N., Lo C.S., Zhao S., Zhao X.P., Chenier I., Yamashita M., Filep J.G., Ingelfinger J.R., Zhang S.L, & Chan J.S. (2021). Deletion of heterogeneous nuclear ribonucleoprotein F in renal tubules downregulates SGLT2 expression and attenuates hyperfiltration and kidney injury in a mouse model of diabetes. Diabetologia, 64(11), 2589-2601.

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Temporal Glucose Dynamics in Hepatocyte Model

Cited 1 time

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After 24 h of FA treatment exposure, media was harvested from each plate, pooled within triplicate, and stored at −80 °C for subsequent quantification of glucose (Autokit Glucose, Wako Diagnostics, Richmond, VA), TG (L-Type Triglyceride M, Wako Diagnostics, Richmond, VA), and BHB (commercial kit, CataChemWell-T, Awareness Technologies, Westport, CT).
After removal of media, cells were harvested for subsequent quantification of cellular TG and DNA as previously described^{16 (link)}. Total cellular TG was normalized to corresponding total DNA within each culture plate prior to averaging within triplicates. Total glucose, BHB, and TG in the pooled media sample were normalized to total DNA averaged across the triplicate.
To characterize the temporal pattern of glucose export in this culture system, a parallel set of isolated hepatocytes were cultured for the serial sampling of media for glucose analysis. After 4 h of incubation following cell seeding, media was refreshed with base media without glucose, Met, and choline chloride and cells were randomly assigned in triplicate to media containing either 0 or 1 mM FA. Cells were cultured for the next 20 h before cell media was refreshed with the same treatments. Over the culture period from 8 to 48 h, 25 µL of media from each triplicate was serially collected and pooled across triplicates every 4 h for subsequent analysis of glucose.

Chandler T.L., Kendall S.J, & White H.M. (2023). Fatty acid challenge shifts cellular energy metabolism in a substrate-specific manner in primary bovine neonatal hepatocytes. Scientific Reports, 13, 15020.

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Plasma Biomarkers Analysis Protocol

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Plasma levels of glucose were measured by a test kit (Autokit Glucose) obtained from Wako Diagnostics (Mountain View, CA). Plasma triglycerides (TGs) and total cholesterol were analyzed by test kits purchased from Fisher-Scientific, Pittsburgh, PA. Plasma HDL cholesterol was determined using MaxDiscovery^™HDL Cholesterol Assay Kit (Bioo Scientific Corporation, Austin, TX). Plasma LDL-C (in mg/dL) was estimated by the Friedewald Equation [21 (link)]: LDL-C = total cholesterol – HDL cholesterol – (total triglycerides/5). Plasma insulin was determined using a mouse insulin ELISA kit (Alpco Diagnostics, Salem, NH). Plasma Monocyte Chemoattractant Protein-1 (MCP-1 or CCL2) and Interleukin (IL)-6 were analyzed by ELISA kits from Life Technologies (Carlsbad, CA). Plasma leptin levels were estimated by an ELISA kit obtained from Crystal Chem, Inc., Downers Grove, IL. Mouse plasma Proprotein Convertase Subtilisin Kexin 9 (PCSK9) was measured using an ELISA kit purchased from R&D Systems (Minneapolis, MN).

Miranda C.L., Elias V.D., Hay J.J., Choi J., Reed R.L, & Stevens J.F. (2016). Xanthohumol improves dysfunctional glucose and lipid metabolism in diet-induced obese C57BL/6J mice. Archives of biochemistry and biophysics, 599, 22-30.

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Epicatechin Reverses Corticosteroid-Induced Insulin Resistance

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Example 6

FIG. 5 depicts the effects of the administration of 10 consecutive days of prednisolone (10 mg/kg/day, SC) on an oral glucose tolerance test. Three groups of male rats were analyzed: 1) control group (saline solution, SC and saline solution by gavage BID); 2) prednisolone (10 mg/kg/day, SC and saline solution by gavage BID) and; 3) prednisolone (10 mg/kg/day, SC) plus epicatechin (Epi; 1 mg/kg/day by gavage BID).

Blood samples were obtained from animals on each group from 0-4 hours after an oral glucose charge (1.25 g/Kg). Glucose levels were determined spectrophotometrically with an autokit glucose (Wako) kit. Abnormal glucose tolerance, indicating insulin resistance associated with corticosteroid therapy, is largely reversed by combining epicatechin with prednisolone (FIG. 5).

US11154546B2. Methods and compositions for treatment of mitochondrial toxicity (2021-10-26). Epirium Bio Inc. [US]. Inventors: Francisco Villarreal [US], Alan Maisel [US], George Schreiner [US], Guillermo M. Ceballos Reyes [MX], Pam Taub [US].

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Metabolic Biomarkers Quantification

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Plasma glucose, insulin, adiponectin, and leptin levels were measured by Autokit Glucose (Wako, Osaka, Japan), Mouse Insulin ELISA KIT (U‐type) (Shibayagi, Japan), Quantikine^® ELISA Mouse Adiponectin/Acrp30 Immunoassay (R&D Systems, Minneapolis, MN, USA), and Quantikine^® ELISA Mouse/Rat Leptin Immunoassay (R&D Systems), respectively. t‐Cho, TG, and NEFA were measured with LabAssay^™ Cholesterol (Wako), LabAssay^™ Triglyceride (Wako), and LabAssay^™ NEFA (Wako), respectively. All assays were performed according to the manufacturers' protocols. Plasma 3‐HB levels were measured by modification of a previously reported method (Hansen & Freier, 1978). Briefly, plasma was added to a reaction buffer, containing 80 mm Tris–HCl (pH 9.5) and 4 mm β‐NAD⁺, and incubated at 37 °C for 5 min. Reactions were initiated by addition of 0.37 U mL⁻¹ 3‐hydroxybutyrate dehydrogenase. Changes in absorbance were measured for 10 min at 340 nm using a SpectraMax Plus384 (Molecular Devices, Sunnyvale, CA, USA).

Fujii N., Narita T., Okita N., Kobayashi M., Furuta Y., Chujo Y., Sakai M., Yamada A., Takeda K., Konishi T., Sudo Y., Shimokawa I, & Higami Y. (2017). Sterol regulatory element‐binding protein‐1c orchestrates metabolic remodeling of white adipose tissue by caloric restriction. Aging Cell, 16(3), 508-517.

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Epicatechin Reverses Glucocorticoid-Induced Insulin Resistance

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Example 6

US10052316B2. Methods and compositions for treatment of mitochondrial toxicity (2018-08-21). None [US], None [US], None [US], None [MX], None [US]. Inventors: Francisco Villarreal [US], Alan Maisel [US], George Schreiner [US], Guillermo M. Ceballos Reyes [MX], Pam Taub [US].

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Glucose Production in Mouse Hepatocytes

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Primary mouse hepatocytes, attached to the collagen plates for 16 h following seeding, were washed twice with PBS and starved in DMEM without glucose for 3 h. Then the cells were washed with PBS, followed by incubation for 3 h in glucose production medium (pH 7.4) consisting of glucose-free DMEM without phenol red, supplemented with 15 mM Hepes and 2 mM L-glutamine, and 10 mM glycerol or 20 mM sodium lactate plus 2 mM sodium pyruvate. After incubation, the medium was collected for glucose measurement (Autokit Glucose; Wako) and the hepatocytes were collected for protein measurement [18 (link)].

Al-Mass A., Poursharifi P., Peyot M.L., Lussier R., Chenier I., Leung Y.H., Ghosh A., Oppong A., Possik E., Mugabo Y., Ahmad R., Sladek R., Murthy Madiraju S.R., Al-Mulla F, & Prentki M. (2022). Hepatic glycerol shunt and glycerol-3-phosphate phosphatase control liver metabolism and glucodetoxification under hyperglycemia. Molecular Metabolism, 66, 101609.

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Effects of Prednisolone and Epicatechin on Glucose Levels

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Example 5

FIG. 4 depicts the effects of the administration of 10 consecutive days of prednisolone (10 mg/kg/day, SC) on glucose levels. Three groups of male rats were analyzed: 1) control group (saline solution, SC and saline solution by gavage BID); 2) prednisolone (10 mg/kg/day, SC and saline solution by gavage BID) and; 3) prednisolone (10 mg/kg/day, SC) plus epicatechin (Epi; 1 mg/kg/day by gavage BID).

Blood samples were obtained from animals on each group after 12 hours fasting, Glucose levels were determined spectrophotometrically with an autokit glucose (Wako) kit. Animals receiving corticosteroid only displayed significant hyperglycemia, those also receiving epicatechin exhibited a marked improvement in their hyperglycemia. (FIG. 4).

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