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Amplex red glucose glucose oxidase kit

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The Amplex Red Glucose/Glucose Oxidase Kit is a fluorometric assay kit used to measure glucose levels. The kit contains the Amplex Red reagent, which reacts with hydrogen peroxide produced by the action of glucose oxidase on glucose, resulting in a fluorescent product that can be measured.

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Lab products found in correlation

Accuri c6 flow cytometer, by BD (1 mentions) Spectramax paradigm multi mode microplate detection platform, by Molecular Devices (1 mentions) Lactate oxidase, by Merck Group (1 mentions) Varioskan multimode microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions) Microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Amplex Red (375 citations) Amplex Red Glucose/Glucose Oxidase Assay Kit (114 citations) Amplex Red kit (70 citations) Glucose oxidase kit (4 citations) Amplex red glucose kit (3 citations) Amplex Red/Glucose Oxidase kit (2 citations)

8 protocols using amplex red glucose glucose oxidase kit

1

Glucose Quantification in Cell Culture

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The
production of glucose was measured by sampling 50 μL of cell
culture medium from the vehicle (negative control, 0.1% DMSO), the
glucose-inducing control (Dex-cAMP)(DC), positive control (Dex-cAMP
with 10 nM insulin)(Ins), and treatment levels (Dex-cAMP with 50 μg/mL
fractions or 10 μM of a single compound 1–9). The conversion of glucose to gluconolactone and hydrogen peroxide
was measured at an absorption of 530 nm and an emission of 590 nm
following the instructions of the Amplex red glucose/glucose oxidase
kit (Life Technologies, Carlsbad, CA) on a fluorescence microplate
reader (BioTek Synergy HA, Sunnyvale, CA). All cellular results are
expressed as the percent of the induced control.
Murray A.F., Palatini K., Komarnytsky S., Gianfagna T.J, & Munafo JP J.r. (2019). Phenylpropanoid Glycerol Glucosides Attenuate Glucose Production in Hepatocytes. ACS Omega, 4(6), 10670-10676.
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2

Quantifying Glucose Uptake in Cell Lines

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U2OS or S180 cells were planted in 6-well plates (2 × 105 cells/well). After incubation over night, RPMI 1640 medium (11 mM glucose) containing 3 % FBS with or without serial concentrations of IS were replaced to the 6-well plates. After cells were treated with IS for 24 h, culture media were collected and diluted 1 : 1000 in water. Glucose in the culture media was quantitated via an Amplex Red Glucose/Glucose Oxidase Kit (Life Technologies, Carlsbad, USA) using a standard curve prepared with serial dilutions of RPMI 1640 (11 mM glucose) into glucose-free RPMI 1640. Fluorescence was read using a Spectra-Max Paradigm Multi-Mode Microplate Detection Platform (Molecular Devices, Sunnyvale, California, United States) at Ex./Em. = 530 nm / 590 nm and normalized to the number of cells in each well counted by BD Accuri™ C6 flow cytometer (Becton & Dickinson Company, Franklin Lakes, NJ). Cell-free medium was used as a background control. The concentration of glucose uptake in each sample was then calculated. Glucose uptake was determined by subtracting the amount of glucose in each sample from the total amount of glucose in cell-free medium (11 mM glucose). In SW1353 cells, the incubation medium was replaced by DMEM medium and glucose uptake was assayed by the same method.
Zhang C., Yang L., Geng Y.D., An F.L., Xia Y.Z., Guo C., Luo J.G., Zhang L.Y., Guo Q.L, & Kong L.Y. (2016). Icariside II, a natural mTOR inhibitor, disrupts aberrant energy homeostasis via suppressing mTORC1-4E-BP1 axis in sarcoma cells. Oncotarget, 7(19), 27819-27837.
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3

Quantifying Lactate Using Amplex Red

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lactate present in culture medium was quantitated using Amplex Red Glucose/Glucose Oxidase kit (Life Technologies) following manufacturer’s instructions, substituting lactate and lactate oxidase (both from Sigma-Aldrich, St. Louis, MO) for glucose and glucose oxidase.
Wyatt E.V., Diaz K., Griffin A.J., Rassmussen J.A., Crane D.D., Jones B.D, & Bosio C.M. (2016). Metabolic reprogramming of host cells by virulent Francisella tularensis for optimal replication and modulation of inflammation. Journal of immunology (Baltimore, Md. : 1950), 196(10), 4227-4236.
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4

Insulin-Mediated Glucose Production in H4IIE Cells

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H4IIE cells in 24-well plates were treated with serial dilutions of human insulin, ExpressTec-ProINS-Tf, HEK-ProINS-Tf or rice-derived Tf (Invitria, Junction City, KS, USA) in serum-free DMEM at 37 °C for 24 h. After the treatment, the dosing solution was replaced with glucose production medium consisting of serum-, glucose- and phenol red-free DMEM supplemented with 2 mM sodium pyruvate and 40 mM sodium DL-lactate, followed by incubation at 37 °C for 3 h. The medium was harvested and glucose concentration of each sample was measured using the Amplex Red Glucose/Glucose Oxidase Kit (Invitrogen). The cells were then lysed with 1N NaOH, and total cellular protein was determined using protein bicinchoninic acid assay for normalization.
Chen Y.S., Zaro J.L., Zhang D., Huang N., Simon A, & Shen W.C. (2018). Characterization and Oral Delivery of Proinsulin-Transferrin Fusion Protein Expressed Using ExpressTec. International Journal of Molecular Sciences, 19(2), 378.
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5

Glucose Quantification during Growth

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Cells were prepared as described, and at FCOD600 = 6 (before glucose exhaustion) and FCOD600 = 30 (after glucose exhaustion), membranes were aseptically removed from the agar and vortexed in 1 mL of PBS. For the no carbon control, samples were taken at FCOD600 = 6 and 2 h post-glucose exhaustion. Glucose was quantified using the Amplex Red Glucose/Glucose Oxidase Kit (Invitrogen, Eugene, OR).
Amato S.M, & Brynildsen M.P. (2014). Nutrient Transitions Are a Source of Persisters in Escherichia coli Biofilms. PLoS ONE, 9(3), e93110.
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6

Quantifying Glucose Uptake in Cells

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Cells were seeded at a density of 2 × 105 cells per well in 6-well cell culture plate. After cells were treated with OA for 10 h, culture media were collected and diluted 1 : 4000 in water. Glucose in the medium was quantitated via an Amplex Red Glucose/ Glucose Oxidase Kit (Invitrogen, Eugene, OR, USA) using a standard curve prepared with serial dilutions of RPMI 1640 (11 mmol/l glucose) into glucose-free RPMI 1640. Fluorescence was read using a Varioskan multimode microplate spectrophotometer (Thermo) at Ex./Em.=530 nm/590 nm, and normalized as the method in the section 'Measurements of lactate generation'. The concentration of glucose uptake in each sample was then calculated. Glucose uptake was determined by subtracting the amount of glucose in each sample from the total amount of glucose in the media without cells (11 mmol/l glucose).
Wei L., Zhou Y., Qiao C., Ni T., Li Z., You Q., Guo Q, & Lu N. (2015). Oroxylin A inhibits glycolysis-dependent proliferation of human breast cancer via promoting SIRT3-mediated SOD2 transcription and HIF1α destabilization. Cell Death & Disease, 6(4), e1714-.
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7

Quantitative Glucose Uptake Assay

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Glucose in the medium was detected by an Amplex Red Glucose/Glucose Oxidase Kit (Invitrogen, Eugene, OR, USA) using a standard curve prepared with serial dilutions of DMEM (11 mmol/L glucose) into glucose-free McCoy’s 5A medium. A microplate spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) was used to read the fluorescence. The glucose uptake level was then calculated.
Yu T., Li L., Liu W., Ya B., Cheng H, & Xin Q. (2019). Silencing of NADPH Oxidase 4 Attenuates Hypoxia Resistance in Neuroblastoma Cells SH-SY5Y by Inhibiting PI3K/Akt-Dependent Glycolysis. Oncology Research, 27(5), 525-532.
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8

Glucose Quantification in Cells

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Glucose in media was measured using Amplex Red Glucose/Glucoseoxidase Kit (Invitrogen) according to the manufacturer’s instructions. Glucose concentration was normalized to total cellular protein.
Groeger M., Matsuo K., Heidary Arash E., Pereira A., Le Guillou D., Pino C., Telles-Silva K.A., Maher J.J., Hsiao E.C, & Willenbring H. (2023). Modeling and therapeutic targeting of inflammation-induced hepatic insulin resistance using human iPSC-derived hepatocytes and macrophages. Nature Communications, 14, 3902.
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