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Oxygraph plus system

Manufactured by Hansatech
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The Oxygraph Plus System is a high-performance laboratory instrument designed to measure oxygen levels in various samples. It provides accurate and reliable data on oxygen consumption or production, which is a crucial parameter in a wide range of scientific and industrial applications.

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Lab products found in correlation

Clark type oxygen electrode, by Hansatech (2 mentions) Cary 100 spectrophotometer, by Agilent Technologies (1 mentions) L galactono 1 4 lactone, by Merck Group (1 mentions) Equine heart cytochrome c, by Merck Group (1 mentions) D arabino 1 4 lactone, by Merck Group (1 mentions) L gulono 1 4 lactone, by Merck Group (1 mentions) Xf analyzer, by Agilent Technologies (1 mentions) Microplate reader, by Tecan (1 mentions) 8453 diode array uv vis spectrophotometer, by Agilent Technologies (1 mentions) Dual pam 100 fluorometer, by Walz (1 mentions) K late kit, by Megazyme (1 mentions)

Spelling variants of this product

Oxygraph (81 citations) Oxygraph system (67 citations) Oxygraph Plus (27 citations) Oxygraph Plus oxygen electrode system (4 citations)

30 protocols using oxygraph plus system

1

Measuring Oxygen Consumption in AAPH

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Solutions of AAPH (100 mM) in phosphate buffer 200 mM, pH 7.0 were incubated in a thermostated cell at 37 °C. O2 consumption was assessed employing an Oxygraph Plus System, Hansatech-Instruments® (Norfolk, UK). The rate of consumption was determined by measuring the decrease in O2 concentration as a function of time. The response of the electrode to O2 was equal in samples containing 0, 30, 60 and 120 mg mL−1 dextran, implying that the initial concentration of O2 was not affected by the presence of the crowding agent.
Fuentes-Lemus E., Reyes J.S., Gamon L.F., López-Alarcón C, & Davies M.J. (2021). Effect of macromolecular crowding on protein oxidation: Consequences on the rate, extent and oxidation pathways. Redox Biology, 48, 102202.
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2

Measuring Yeast Cell Respiration Rates

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Oxygen consumption rate (ORC) of intact yeast cells was analyzed via the
polarographic method in a closed 500 μL chamber equipped with a micro
Clark-type oxygen electrode (Oxygraph Plus System, Hansatech Instruments) at
30°C. Yeast cells were grown in synthetic complete medium until log
phase. Cells were mixed in fresh medium with a cell density normalized to a
protein concentration of 0.5 mg/mL followed by the measurement of respiration.
KCN (0.2 mM) was added at the end of the experiment to inhibit cytochrome c
oxidase, to correct for cytochrome c oxidase-independent oxygen utilization.
Oxygen consumption was recorded on a computer and analyzed with Oxygraph plus
software. Oxygen consumption rate (ORC) is defined as consumed O2(nmol)/min/total protein (mg).
Lou W., Ting H.C., Reynolds C.A., Tyurina Y.Y., Tyurin V.A., Li Y., Ji J., Yu W., Liang Z., Stoyanovsky D.A., Anthonymuthu T.S., Frasso M.A., Wipf P., Greenberger J.S., Bayir H., Kagan V.E, & Greenberg M.L. (2018). Genetic re-engineering of polyunsaturated phospholipid profile of Saccharomyces cerevisiae identifies a novel role for Cld1 in mitigating the effects of cardiolipin peroxidation. Biochimica et biophysica acta. Molecular and cell biology of lipids, 1863(10), 1354-1368.
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3

Daphnid Oxygen Consumption Assay

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Oxygen consumption was determined with Oxygraph plus system (Hansatech Instruments) according to previously published methods (Soucek 2006 (link); Soucek et al. 2010 ) with some modifications. 10 daphnids were transfered to the oxygraph electrode chamber containing 1 ml of medium with appropriate concentration of the antibiotic and the oxygen consumption was measured for 30 min. The oxygen was detected by an electrode mounted at the bottom of the chamber and the signal from the electrode was transferred to a computer. Oxygen consumption rate was calculated by computer software, O2view version 2.09. The experiment was done in duplicate.
Bownik A., Ślaska B., Bochra J., Gumieniak K, & Gałek K. (2019). Procaine penicillin alters swimming behaviour and physiological parameters ofDaphnia magna. Environmental Science and Pollution Research International, 26(18), 18662-18673.
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4

Polarographic Measurement of Horsehair Worm Oxygen Consumption

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The oxygen consumption in the horsehair worm tissue homogenate suspensions was determined polarographically using an Oxygraph Plus System containing the thermostatically controlled chamber with a Clark-type oxygen electrode (Hansatech Instruments, England) connected to PC. The digital signals incoming from Oxygraph System to PC were recorded with the software ‘O2View’ supplied by the manufacturer. All the measurements were performed in the sample volume of 250 μl under vigorous stirring at 37°C; the incubation medium contained 20 mM Mes/Tris (pH 6.0), 2.5 mM MgSO4, 0.2 mM EGTA, antibiotic alamethicin 50 μg/ml and 200 μM NADH. The oxygen consumption was started by the addition of tissue homogenate sample. The measurements were performed in three biological replicas, i.e. with three different tissue suspensions.
Mikhailov K.V., Efeykin B.D., Panchin A.Y., Knorre D.A., Logacheva M.D., Penin A.A., Muntyan M.S., Nikitin M.A., Popova O.V., Zanegina O.N., Vyssokikh M.Y., Spiridonov S.E., Aleoshin V.V, & Panchin Y.V. (2019). Coding palindromes in mitochondrial genes of Nematomorpha. Nucleic Acids Research, 47(13), 6858-6870.
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5

Oxygen Consumption Assay for FIH Activity

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An Oxygraph Plus System (Hansatech) was used to monitor the amount of oxygen consumed by the enzyme. This oxygen sensor contains a central reaction vessel surrounded by a water jacket, with a Clark-type electrode disc at the bottom of the reaction vessel. The electrode disc utilized a KCl bridge and a PTFE membrane that is selectively permeable to oxygen molecules. A new membrane was prepared and calibrated each day with air saturated water and dithionite. 400 μL reactions were equilibrated at atmospheric O2 in the reaction vessel at 37 °C until a stable baseline was achieved. Assays were initiated with cold purified FIH (10 μM) using a Hamilton gas-tight syringe. The rate of O2 consumption was monitored over time until the rate of O2 consumption resembled the baseline slope. Assays contained 100 μM αKG, 50 μM FeSO4, 80 μM CTAD and 10 μM FIH in 50 mM HEPES pH 7.00, and 50 μM ascorbate. Controls were performed to determine the optimal amount of ascorbate that could be used to minimize baseline slope while retaining maximal FIH activity. 5 μL of each reaction was then quenched in 20 μL of matrix (3,5-dimethoxy-4-hydroxycinnamic acid saturated in 75% ACN/0.2% TFA) and analyzed by MALDI-TOF-MS (Ultra-flex, Bruker) monitoring the +1 charge state of the substrate (CAD) and the product (hydroxylated CAD).
Iyer S.R., Chaplin V.D., Knapp M.J, & Solomon E.I. (2018). O2 Activation by Nonheme FeII α-Ketoglutarate Dependent Enzyme Variants: Elucidating the Role of the Facial Triad Carboxylate in FIH.. Journal of the American Chemical Society, 140(37), 11777-11783.
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6

Yeast Oxygen Consumption Analysis

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The oxygen consumption rate (ORC) of intact yeast cells was analyzed via the polarographic method in a closed 500 μL chamber equipped with a micro Clark-type oxygen electrode (Oxygraph Plus System, Hansatech Instruments) at 30 °C. Yeast cells were grown in synthetic YPD media supplemented with oleic acid, linoleic acid, or arachidonic acid. Cells were harvested during the stationary phase. Cells were mixed in fresh media using a final protein concentration of 0.5 mg mL−1 following measurements of respiration. KCN (0.2 mM) was added at the end of the experiment to inhibit cytochrome c oxidase to correct for cytochrome c oxidase-independent oxygen utilization. Oxygen consumption was recorded on a computer and analyzed with the Oxygraph plus software. Oxygen consumption rate (ORC) is defined as consumed O2 (nmol)/min/total protein (mg).
Tyurina Y.Y., Lou W., Qu F., Tyurin V.A., Mohammadyani D., Liu J., Hüttemann M., Frasso M.A., Wipf P., Bayir H., Greenberg M.L, & Kagan V.E. (2016). Lipidomics Characterization of Biosynthetic and Remodeling Pathways of Cardiolipins in Genetically and Nutritionally Manipulated Yeast Cells. ACS chemical biology, 12(1), 265-281.
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7

Measurement of Cellular Oxygen Consumption

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In all, 5.0 × 106 HepaRG cells were washed with PBS and resuspended in 10 mM KH2PO4, 27 mM KCl, 1 mM MgCl2, 40 mM HEPES, 0.5 mM EGTA buffer (pH 7.1), and assayed for O2 consumption by the Oxygraph Plus System (Hansatech Instruments) at 37 °C under continuous stirring. Oligomycin (8 µg/ml) was added followed by the addition of valinomycin (2 µg/ml) after 5 min. The rates of oxygen consumption were corrected for 3 mM KCN-insensitive respiration and normalized to the cell number. Each experiment was repeated in triplicate.
Bellanti F., di Bello G., Iannelli G., Pannone G., Pedicillo M.C., Boulter L., Lu W.Y., Tamborra R., Villani R., Vendemiale G., Forbes S.J, & Serviddio G. (2021). Inhibition of nuclear factor (erythroid-derived 2)-like 2 promotes hepatic progenitor cell activation and differentiation. NPJ Regenerative Medicine, 6, 28.
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8

Oxygen Consumption Rate Measurement

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Cells were grown to the PDS phase in SC medium, and the oxygen consumption rate was measured for 1 × 107 cells in PBS, using a Clark-type oxygen electrode coupled to an Oxygraph plus system (Hansatech, Norfolk, UK). Data were analyzed using the OxyTrace + software v1.0.48.
Martins T.S., Costa R.S., Vilaça R., Lemos C., Teixeira V., Pereira C, & Costa V. (2023). Iron Limitation Restores Autophagy and Increases Lifespan in the Yeast Model of Niemann–Pick Type C1. International Journal of Molecular Sciences, 24(7), 6221.
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9

Cytochrome c Dehydrogenase Kinetics

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L-galactono-1,4-lactone, L-gulono-1,4-lactone, and D-arabino-1,4-lactone were obtained from Sigma-Aldrich. Dehydrogenase activities were evaluated by monitoring the reduction of 50 μM equine cytochrome c at 550 nm (ε515 = 21 mM−1 cm−1) at 25 °C using a Cary100 spectrophotometer (Agilent). Equine heart cytochrome c (Sigma-Aldrich) was solubilized in storage buffer. Oxidase activities were assayed using an Oxygraph Plus system (Hansatech Instruments Ltd.). Initial rates were determined by analyzing the initial part of the oxygen depletion curve. In all assays, the reactions were initiated by adding the enzymes (20 nM GalDH, 1.0 μM GULO) to the system. Data were processed in GraphPad Prism and fitted using a standard Michaelis Menten formula resulting in KM and kcat values. All experiments were performed in duplicates.
Boverio A., Jamil N., Mannucci B., Mascotti M.L., Fraaije M.W, & Mattevi A. (2024). Structure, mechanism, and evolution of the last step in vitamin C biosynthesis. Nature Communications, 15, 4158.
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10

Measuring Cellular Oxygen Consumption

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OCR was assessed using a Clark-type electrode in samples prepared from cultures on day 1. Briefly, cells were collected by centrifugation, and the cell pellet was resuspended in 1 ml of 2% glucose-containing YPD medium. Then, oxygen consumption was measured in these cells using an Oxygraph Plus system (Hansatech instruments, UK) at 30°C.
Kwon Y.Y., Choi K.M., Cho C, & Lee C.K. (2015). Mitochondrial Efficiency-Dependent Viability of Saccharomyces cerevisiae Mutants Carrying Individual Electron Transport Chain Component Deletions. Molecules and Cells, 38(12), 1054-1063.
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