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Ros glo h2o2

Manufactured by Promega
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Sourced in France, United States

The ROS-Glo H2O2 is a luminescent assay that measures hydrogen peroxide (H2O2) levels in biological samples. It provides a sensitive and quantitative way to detect and monitor H2O2 production in cells or tissues. The assay employs a proprietary luminogenic substrate that reacts with H2O2 to generate a stable luminescent signal proportional to the H2O2 concentration.

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Lab products found in correlation

Celltiter glo 2, by Promega (3 mentions) Caspase glo 3 7, by Promega (3 mentions) Celltiter glo, by Promega (2 mentions) Calcein am, by Thermo Fisher Scientific (2 mentions) Cyto id autophagy detection kit 2, by Enzo Life Sciences (1 mentions) Prestoblue, by Thermo Fisher Scientific (1 mentions) Glucose colorimetric fluorometric assay kit, by Abcam (1 mentions) Jc 10, by Enzo Life Sciences (1 mentions) Human cardiac troponin 1 elisa kit, by RayBiotech (1 mentions) Cytation 5, by Agilent Technologies (1 mentions) Luminoskan microplate luminometer, by Thermo Fisher Scientific (1 mentions) Mtesr1, by STEMCELL (1 mentions) Glomax multi detection system, by Promega (1 mentions) Gsh glo glutathione assay kit, by Promega (1 mentions) Nad nadh glo, by Promega (1 mentions) Menadione, by MedChemExpress (1 mentions) Staurosporine, by MedChemExpress (1 mentions) Varioskan lux multimode reader, by Thermo Fisher Scientific (1 mentions) Prism 7, by GraphPad (1 mentions) Omega plate reader, by BMG Labtech (1 mentions)

Spelling variants of this product

ROS-Glo™ H2O2 Assay (148 citations) ROS-Glo H2O2 assay kit (55 citations) ROS-Glo (10 citations) ROS-GloTM H2O2 assay kit (8 citations) ROS-GloTM H2O2 assay (8 citations) ROS-Glo H2O2 kit (6 citations) ROS-Glo H2O2 luminescence-based detection system (4 citations) ROS-Glo H2O2 luminescence kit (2 citations) ROS-Glo™ H2O2 detection solution (2 citations) ROS-Glo™ H2O2 luminescence assay (2 citations)

13 protocols using ros glo h2o2

1

Plate-based Assays for iPSC-Derived Cardiomyocytes

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Human iPSC-CMs plated on 96-well plates were subjected to plate-based assays after 7 days of drug treatment using a Cytation 5 plate reader/imager (BioTek Instruments). PrestoBlue (Life Technologies) was used for cellular viability assay, CYTO-ID Autophagy detection kit 2.0 (Enzo Life Sciences) was used for autophagy assay, JC-10 (Enzo Life Sciences) was used for mitochondrial membrane potential measurement, ROS-Glo H2O2 (Promega) was used for ROS detection, and Cell Titer Glo (Promega) was used for ATP content assay. For measurement of troponin I and glucose levels in cell culture media, day 6–7 media were collected during 7 days of drug treatment and centrifuged to remove dead cells. Troponin I and glucose concentrations in the supernatant were measured by using the Glucose Colorimetric/Fluorometric Assay Kit (BioVision) and Human Cardiac Troponin I ELISA kit (RayBiotech), respectively. All assays were performed according to the manufacturer’s instructions.
Kitani T., Ong S.G., Lam C.K., Rhee J.W., Zhang J.Z., Oikonomopoulos A., Ma N., Tian L., Lee J., Telli M.L., Witteles R.M., Sharma A., Sayed N, & Wu J.C. (2019). Human Induced Pluripotent Stem Cell Model of Trastuzumab-Induced Cardiac Dysfunction in Breast Cancer Patients. Circulation, 139(21), 2451-2465.
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2

Measurement of ROS and H₂O₂ in Egg Yolks

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The contents of ROS, hydrogen peroxide (H₂O₂) from 10 egg yolks per group in each experiment were measured by Ros-Glo H2O2 assay (Promega Corporation, Charbonnières-les-Bains, France). In each analysis, one gram of each egg yolk for each group was precisely analysed following the manufacturer’s protocol.
Barbe A., Mellouk N., Ramé C., Grandhaye J., Anger K., Chahnamian M., Ganier P., Brionne A., Riva A., Froment P, & Dupont J. (2020). A grape seed extract maternal dietary supplementation improves egg quality and reduces ovarian steroidogenesis without affecting fertility parameters in reproductive hens. PLoS ONE, 15(5), e0233169.
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3

ROS Assay of H9 ESC Lines

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Human H9 ESC lines with normal and abnormal karyotype (trisomy of chromosome 12) were harvested with Accutase, counted and plated on Matrigel-coated 96-well plates in mTeSR1 (Stem Cell Technologies). ROS assay (ROS-Glo™ H2O2, Promega) was carried out to according to the kit manual following the Cell-Based assay. Also matrigel-coated wells with mTeSR1 and without cells were included in the measurements. The luminescent signal was measured 2–3 hours after incubation with H2O2 substrate and after 20 min incubation with ROS-GloTM reagent with Luminoskan Microplate Luminometer (Thermo Scientific). Three biological and four biological replicates were carried out for karyotypically normal and abnormal cells, respectively, and the signals for each replicate were measured from at least 9 wells.
Konki M., Pasumarthy K., Malonzo M., Sainio A., Valensisi C., Söderström M., Emani M.R., Stubb A., Närvä E., Ghimire B., Laiho A., Järveläinen H., Lahesmaa R., Lähdesmäki H., Hawkins R.D, & Lund R.J. (2016). Epigenetic Silencing of the Key Antioxidant Enzyme Catalase in Karyotypically Abnormal Human Pluripotent Stem Cells. Scientific Reports, 6, 22190.
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4

iPSC-ECs Toxicity Screening

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iPSC-ECs plated on 96-well plates were subjected to plate-based assays after serum (15%) or uremic toxin mixture treatments. Assays using CellTiter-Glo 2.0, ROS-Glo H2O2, and Caspase-Glo 3/7 (Promega, Madison, WI) were performed following the manufacturer's instruction and the luminescence signal was recorded on the GloMax-Multi detection system (Promega). Cells in the original sample plate were kept for measuring the total cell number by calcein-AM (Thermo Fisher Scientific) to allow normalization.
Jang H.R., Cho H.J., Zhou Y., Shao N.Y., Lee K., Le H.H., Jeon J., Lee J.E., Huh W., Ong S.G., Lee W.H, & Kim Y.G. (2021). Modeling Uremic Vasculopathy With Induced Pluripotent Stem Cell-Derived Endothelial Cells as a Drug Screening System. Frontiers in Cell and Developmental Biology, 8, 618796.
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5

Oxidative Stress Measurement Assay

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The levels of H2O2 (ROS) and glutathione (antioxidant) were measured using the ROS-Glo™ H2O2 and GSH-Glo™ Glutathione assay kits (Promega, Wisconsin, USA) respectively following the manufacturer’s instructions and the results were read using the GloMax® Explorer machine (Promega, Wisconsin, USA).
Mohamed L., Chakraborty S., ArulJothi K.N., Mabasa L., Sayah K., Costa-Lotufo L.V., Jardine A, & Prince S. (2020). Galenia africana plant extract exhibits cytotoxicity in breast cancer cells by inducing multiple programmed cell death pathways. Saudi Pharmaceutical Journal : SPJ, 28(10), 1155-1165.
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6

Quantifying Cellular Metabolites

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ATP (CellTiter-Glo® 2.0), hydrogen peroxide (H2O2) (ROS-Glo™ H2O2), and NAD+ (NAD/NADH-Glo™) levels were assayed at 2 h after treatments according to the protocol (Promega, Madison, WI).
Jiang L., Liu Y., Su X., Wang J., Zhao Y., Tumbath S., Kilgore J.A., Williams N.S., Chen Y., Wang X., Mendonca M.S., Lu T., Fu Y.X, & Huang X. (2022). KP372-1-Induced AKT Hyperactivation Blocks DNA Repair to Synergize With PARP Inhibitor Rucaparib via Inhibiting FOXO3a/GADD45α Pathway. Frontiers in Oncology, 12, 976292.
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7

Measuring Cellular ROS Levels

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To measure ROS production, the ROS-Glo H2O2 (Promega, Madison, WI, USA) assay was conducted according to the manufacturer’s protocol, which measures the most stable ROS-H2O2 level. Cells were plated at a density of 5 × 103/70 μL medium in white-walled opaque bottom 96-well plates and incubated overnight (37 °C, 5% CO2). TQ or medium (10 μL) was added to treated or control wells, respectively, for an extra 6 h and 24 h. Then, 20 μL H2O2 substrate was added to each well either simultaneously with TQ in the 6 h group or after 18 h in the 24 h group. Finally, 100 μL ROS-Glo solution was added to each well, and after 20 min, relative luminescence was measured in a plate reader (PerkinElmer Life and Analytical Sciences, Buckinghamshire, UK).
Al-Hayali M., Garces A., Stocks M., Collins H, & Bradshaw T.D. (2021). Concurrent Reactive Oxygen Species Generation and Aneuploidy Induction Contribute to Thymoquinone Anticancer Activity. Molecules, 26(17), 5136.
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8

Dose-dependent Cytotoxicity and Apoptosis Assay

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To measure cell viability after 72 h of doxorubicin (0.01-100 μM) treatment, CellTiter-Glo 2.0 (Promega) was used per manufacturer’s instructions. Luminescence was measured using a VarioSkan Lux Multi-Mode Reader (Thermo Scientific) with an integration time of 0.25 sec. 10 μM staurosporine (MedChemExpress) was used as a positive control. After 24 h of doxorubicin (0.01-100 μM) treatment, apoptosis and ROS was measured using Caspase 3/7-Glo and ROS-Glo H2O2 (both Promega) respectively according to manufacturer’s instructions with an integration time of 1 sec. 10 μM staurosporine and 50 μM menadione (both MedChemExpress) was used as a positive control, respectively. Data were analyzed using Prism 7.0 software (GraphPad) using standard dose-response guidelines.
Magdy T., Jiang Z., Jouni M., Fonoudi H., Lyra-Leite D., Jung G., Romero-Tejeda M., Kuo H.H., Fetterman K.A., Gharib M., Burmeister B.T., Zhao M., Sapkota Y., Ross C.J., Carleton B.C., Bernstein D, & Burridge P.W. (2021). RARG variant predictive of doxorubicin-induced cardiotoxicity identifies a cardioprotective therapy. Cell stem cell.
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9

Quantification of Cellular ROS Levels

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Reactive oxygen species (ROS) levels present in treated and non-treated HK-2 cells were assayed using ROS-Glo H 2 O 2 (Promega), following the manufacturer's instructions and data were normalized to cell viability as previously described (Mossoba et al., 2016a (Mossoba et al., , 2016b)) . Cells plated in white-wall, clear-bottom plates were analyzed for luminescence emission using the Omega plate reader (BMG Labtech). Experiments were independently performed at least three times.
Mossoba M.E., Mapa M.S.T., Araujo M., Zhao Y., Flannery B., Flynn T., Sprando J., Wiesenfeld P, & Sprando R.L. (2020). Long-term in vitro effects of exposing the human HK-2 proximal tubule cell line to 3-monochloropropane-1,2-diol. The Journal of toxicological sciences, 45(1).
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10

Plate-based Assays for iPSC-CM

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Human iPSC-CMs plated on 96-well plates were subjected to plate-based assays after 5 days of iron ± drug treatment using a Cytation 5 plate reader/imager (BioTek). ROS-Glo H2O2 (Promega) was used for ROS detection, JC-10 (Abcam) was used for mitochondrial membrane potential, and Cell Titer Glo (Promega) was used for cell viability assay. All assays were performed according to the manufacturer’s instructions.
Rhee J.W., Yi H., Thomas D., Lam C.K., Belbachir N., Tian L., Qin X., Malisa J., Lau E., Paik D.T., Kim Y., Choi B.S., Sayed N., Sallam K., Liao R, & Wu J.C. (2020). Modeling Secondary Iron Overload Cardiomyopathy with Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Cell reports, 32(2), 107886.
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