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ROS-ID® Total ROS detection kit

Widely cited kit to measure global levels of ROS in live cells
ENZ-51011 1 Kit 220.00 USD
Do you need bulk/larger quantities?
  • Directly monitors global levels of reactive oxygen species (ROS), but not superoxide in live cells
  • High sensitivity, specificity and accuracy for live cell studies
  • Compatible with major components of tissue culture media (phenol red, FBS and BSA)
  • Not designed to detect superoxide, reactive chlorine or bromine species
  • Validated on microscopy, flow cytometry, and microplates
Enzo Life Sciences ROS-ID® Total ROS detection kit includes the Oxidative Stress Detection Reagent (Green, Ex/Em 490/525 nm) to directly monitor real time reactive oxygen and/or nitrogen species (ROS/RNS) production in live cells. This non-fluorescent, cell-permeable detection probe reacts directly with a wide range of reactive species (hydrogen peroxide, peroxynitrite and hydroxyl radicals) yielding a green fluorescent product. Upon staining, the fluorescent product generated can be visualized using a wide-field fluorescence microscope equipped with standard green filter (490/525 nm), or cytometrically using any flow cytometer equipped with a blue (488 nm) laser.
ROS-ID® Total ROS detection kit Flow Cytometry
Figure 1. Jurkat cells were induced with 100µM pyocyanin (general ROS inducer, panel A), or 1 µM of t-butyl-hydroperoxide (peroxide inducer, panel B), stained with Total ROS Detection Reagent and analyzed using flow cytometry. Untreated cells were used as a control. Cell debris were ungated. The numbers in the inserts reflect the mean green fluorescence of the control and treated cells.
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ROS-ID® Total ROS detection kit Flow Cytometry

Product Details

Alternative Name:Reactive oxygen species
Applications:Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS
Application Notes:This kit is designed to directly monitor real time reactive oxygen and/or nitrogen species (ROS/RNS) production in live cells using fluorescence microscopy and/or flow cytometry.
Quality Control:A sample from each lot of ROS-ID® Total ROS detection kit is used to stain HeLa cells using the procedures described in the user manual. The stained cells are analyzed using a wide-field fluorescence microscope equipped with standard green filter (490/525 nm).
The following results are obtained: ROS positive control samples induced with Pyocyanin exhibit bright green fluorescence in the cytoplasm. Cells pretreated with the ROS inhibitor don’t demonstrate any green fluorescence signal upon induction.
Quantity:200 fluorescence microscopy assays or 50 flow cytometry assays.
Use/Stability:With proper storage, the kit components are stable up to the date noted on the product label. Store kit at -20°C in a non-frost free freezer, or -80°C for longer term storage.
Handling:Protect from light. Avoid freeze/thaw cycles.
Shipping:Dry Ice
Short Term Storage:-20°C
Long Term Storage:-80°C
Contents:Oxidative Stress Detection Reagent (Green), 300 nmoles 
ROS Inducer (Pyocyanin), 1 µmole
ROS Inhibitor (N-acetyl-L-cysteine), 2 x 10 mg
Wash Buffer Salts, 1 pack
Technical Info/Product Notes:The ROS-ID® Total ROS detection kit is a member of the CELLESTIAL® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications. CELLESTIAL® reagents and kits are optimal for use in demanding imaging applications, such as confocal microscopy, flow cytometry and HCS, where consistency and reproducibility are required.

Application Note:
Image-Based Analysis of a Human Neurosphere Stem Cell Model for the Evaluation of Potential Neurotoxicants
Regulatory Status:RUO - Research Use Only

Product Literature References

A HIF-1α inhibitor combined with palmitic acid and L-carnitine treatment can prevent the fat metabolic reprogramming under hypoxia and induce apoptosis in hepatocellular carcinoma cells: S. Matsufuji, et al.; Cancer Metab. 11, 25 (2023), Abstract;
Allele-dependent interaction of LRRK2 and NOD2 in leprosy: M.D. Sauer, et al.; PLoS Pathog. 19, e1011260 (2023), Abstract;
Chemotherapy-induced tumor immunogenicity is mediated in part by megakaryocyte-erythroid progenitors: A. Vorontsova, et al.; Oncogene 42, 771 (2023), Abstract;
Combination Therapies Targeting Apoptosis in Paediatric AML: Understanding the Molecular Mechanisms of AML Treatments Using Phosphoproteomics: A.A. Ali, et al.; Int. J. Mol. Sci. 24, 5717 (2023), Abstract;
Targeting Unc51-like Autophagy Activating Kinase 1 (ULK1) Overcomes Adaptive Drug Resistance in Acute Myelogenous Leukemia: S. Bhattacharya, et al.; Mol. Cancer Res. 10.1158, 1541 (2023), Abstract;
FOXM1 network in association with TREM1 suppression regulates NET formation in diabetic foot ulcers: A.P. Sawaya, et al.; EMBO Rep. 23, e54558 (2022), Abstract;
Increased clearance of non-biodegradable polystyrene nanoplastics by exocytosis through inhibition of retrograde intracellular transport: S.W. Han, et al.; J. Hazard. Mater. 439, 129576 (2022), Abstract;
Metabolic stress induces GD2+ cancer stem cell-like phenotype in triple-negative breast cancer: A. Jaggupilli, et al.; Br. J. Cancer 126, 615 (2022), Abstract;
Tension exerted on cells by magnetic nanoparticles regulates differentiation of human mesenchymal stem cells: S. Cho, et al.; Biomater. Adv. 139, 213028 (2022), Abstract;
Age-dependent instability of mature neuronal fate in induced neurons from Alzheimer’s patients: J. Mertens, et al.; Cell Stem Cell 28, 1533 (2021), Abstract;
Antitumor effects of low-dose tipifarnib on the mTOR signaling pathway and reactive oxygen species production in HIF-1α-expressing gastric cancer cells: N. Egawa, et al.; FEBS Open Bio. 11, 1465 (2021), Application(s): Flow Cytometry; carcinoma cell lines, Abstract; Full Text
Complement C5a promotes antigen cross-presentation by Peyer's patch monocyte-derived dendritic cells and drives a protective CD8 + T cell response: S.H. Kim, et al.; Cell Rep. 35, 108995 (2021), Abstract;
Dihydroisotanshinone I induced ferroptosis and apoptosis of lung cancer cells: C.Y. Wu, et al.; Biomed. Pharmacother. 139, 111585 (2021), Abstract;
Lipocalin 2 regulates expression of MHC class I molecules in Mycobacterium tuberculosis-infected dendritic cells via ROS production: J.A. Choi, et al.; Cell Biosci. 11, 175 (2021), Abstract;
The Cell Protective Effect of Adenine on Hypoxia–Reoxygenation Injury through PPAR Delta Activation: J.G. Leu, et al.; Life (Basel) 11, 1408 (2021), Abstract;
BETP degradation simultaneously targets acute myelogenous leukemic stem cells and the microenvironment: S. Piya, et al.; J. Clin. Invest. 129, 1878 (2019), Application(s): Flow cytometry using AML cells, Abstract;
Copper oxide nanoparticles inhibit pancreatic tumor growth primarily by targeting tumor initiating cells: M. Benquiqui, et al.; Sci. Rep. 9, 12613 (2019), Abstract; Full Text
Mieap-induced accumulation of lysosomes within mitochondria (MALM) regulates gastric cancer cell invasion under hypoxia by suppressing reactive oxygen species accumulation: K. Okuyama, et al.; Sci. Rep. 9, 2822 (2019), Abstract; Full Text
PUMILIO hyperactivity drives premature aging of Norad-deficient mice: F. Kopp, et al.; Elife 8, e42650 (2019), Application(s): Flow cytometry using immortalized MEFs and HCT116 cells, Abstract; Full Text
Rational combination with PDK1 inhibition overcomes cetuximab resistance in head and neck squamous cell carcinoma: H. Lu, et al.; JCI Insight 4, e131106 (2019), Abstract; Full Text
Activation of AMPK inhibits inflammatory response during hypoxia and reoxygenation through modulating JNK-mediated NF-κB pathway: X. Chen, et al.; Metabolism 83, 256 (2018), Abstract;
Adipocyte-activated oxidative and ER stress pathways promote tumor survival in bone via upregulation of heme oxygenase 1 and survivin: M.K. Herroon, et al.; Sci. Rep. 8, 40 (2018), Application(s): Confocal microscopy with ARCaP(M) and PC3 cells, Abstract; Full Text
Carbonyl reductase 1 is a new target to improve the effect of radiotherapy on head and neck squamous cell carcinoma: M. Yun, et al.; J. Exp. Clin. Cancer Res. 37, 264 (2018), Abstract; Full Text
Novel β-phenylacrylic acid derivatives exert anti-cancer activity by inducing Src-mediated apoptosis in wild-type KRAS colon cancer: S.J. Kim, et al.; Cell Death Dis. 9, 877 (2018), Abstract; Full Text
AP1G1 is involved in cetuximab-mediated downregulation of ASCT2-EGFR complex and sensitization of human head and neck squamous cell carcinoma cells to ROS-induced apoptosis: X. Tao, et al.; Cancer Lett. 408, 33 (2017), Application(s): Ros Detection, Abstract;
HCV-induced oxidative stress by inhibition of Nrf2 triggers autophagy and favors release of viral particles: R. Medvedev, et al.; Free Radic. Biol. Med. 110, 300 (2017), Application(s): Fluorescence microplate reader using HuH7.5.1 cells, Abstract;
Induction of mitophagy-mediated antitumor activity with folate-appended methyl-β-cyclodextrin: K. Kameyama, et al.; Int. J. Nanomedicine 12, 3433 (2017), Application(s): Fluorescence microscopy using A549 and KB cells, Abstract;
Low dose radiation prevents doxorubicin-induced cardiotoxicity: X. Jiang, et al.; Oncotarget 9, 332 (2017), Abstract; Full Text
Modulation of alveolar macrophage innate response in proinflammatory-, pro-oxidant-, and infection- models by mint extract and chemical constituents: Role of MAPKs: N. Yadav & H. Chandra; Immunobiology 223, 49 (2017), Abstract;
Salvianolic Acid A Protects H9c2 Cells from Arsenic Trioxide-Induced Injury via Inhibition of the MAPK Signaling Pathway: J.Y. Zhang, et al.; Cell. Physiol. Biochem. 41, 1957 (2017), Abstract; Full Text
Sustained O-GlcNAcylation reprograms mitochondrial function to regulate energy metabolism: E.P. Tan, et al.; J. Biol. Chem. 292, 14940 (2017), Application(s): Fluorescence microplate reader using NT2 and SH-SY5Y cells, Abstract; Full Text
Anti-inflammatory and antioxidant activity of the traditional herbal formula Gwakhyangjeonggi-san via enhancement of heme oxygenase-1 expression in RAW264.7 macrophages: S.J. Jeong, et al.; Mol. Med. Rep. 13, 4365 (2016), Abstract;
ASCT2 (SLC1A5) is an EGFR-associated protein that can be co-targeted by cetuximab to sensitize cancer cells to ROS-induced apoptosis: H. Lu, et al.; Cancer Lett. 381, 23 (2016), Application(s): Intracellular ROS detection, Abstract;
E-cigarette aerosol exposure induces reactive oxygen species, DNA damage, and cell death in vascular endothelial cells: C. Anderson, et al.; Toxicol. Sci. 154, 332 (2016), Abstract;
Equine Metabolic Syndrome Affects Viability, Senescence, and Stress Factors of Equine Adipose-Derived Mesenchymal Stromal Stem Cells: New Insight into EqASCs Isolated from EMS Horses in the Context of Their Aging: K. Marycz, et al.; Oxid. Med. Cell. Longev. 2016, Article ID 4710326 (2016), Application(s): Fluorescence staining of ROS, Abstract; Full Text
Estrogen Protects the Female Heart from Ischemia/Reperfusion Injury through Manganese Superoxide Dismutase Phosphorylation by Mitochondrial p38β at Threonine 79 and Serine 106: T. Luo, et al.; PLoS One 11, e0167761 (2016), Abstract;
Infection-Mediated Priming of Phagocytes Protects against Lethal Secondary Aspergillus fumigatus Challenge: A. Savers, et al.; PLoS One 11, e0153829 (2016), Application(s): Flow cytometry, Abstract; Full Text
Overcoming cisplatin resistance of ovarian cancer cells by targeting HIF-1-regulated cancer metabolism: Z. Ai, et al.; Cancer Lett. 373, 36 (2016), Application(s): Detection of intracellular ROS, Abstract; Full Text
Anti-adipogenic and antioxidant effects of the traditional Korean herbal formula samchulgeonbi-tang: an in vitro study: S. Yoo, et al.; Int. J. Clin. Exp. Med. 8, 8698 (2015), Application(s): Detection of ROS generation, Abstract; Full Text
C-reactive protein stimulates RAGE expression in human coronary artery endothelial cells in vitro via ROS generation and ERK/NF-κB activation: Y. Zhong, et al.; Acta Pharmacol. Sin. 36, 440 (2015), Application(s): Flow Cytometry, Abstract;
Comparative safety evaluation of silica-based particles: H. Kettiger, et al.; Toxicol. in Vitro 30, 355 (2015), Application(s): Measurement of oxidative stress in microplate reader, Abstract;
Inhibitory effects of oleoylethanolamide (OEA) on H2O2-induced human umbilical vein endothelial cell (HUVEC) injury and apolipoprotein E knockout (ApoE-/-) atherosclerotic mice: L. Ma, et al.; Int. J. Clin. Exp. Pathol. 8, 6301 (2015), Application(s): OEA on intracellular ROS levels, Abstract; Full Text
The Cytoprotective Effects of E-α-(4-Methoxyphenyl)-2’,3,4,4'-Tetramethoxychalcone (E-α-p-OMe-C6H4-TMC)—A Novel and Non-Cytotoxic HO-1 Inducer: K.B. Kaufmann, et al.; PLoS One 10, e0142932 (2015), Application(s): Reactive oxygen species detection in RAW264.7 cells, Abstract;
Transfer hydrogenation catalysis in cells as a new approach to anticancer drug design: J. J. Soldevila-Barreda, et al.; Nat. Commun. 6, 6582 (2015), Application(s): Flow Cytometry, Abstract; Full Text
Cannabidiol protects liver from binge alcohol-induced steatosis by mechanisms including inhibition of oxidative stress and increase in autophagy: L. Yang, et al.; Free Radic. Biol. Med. 68C, 260 (2014), Application(s): Measurement of ROS by flow cytometry, Abstract;
Natural compound Alternol induces oxidative stress-dependent apoptotic cell death preferentially in prostate cancer cells: Y. Tang, et al.; Mol. Cancer Ther. 13, 1526 (2014), Abstract;
Maternal obesity programs offspring nonalcoholic fatty liver disease by innate immune dysfunction in mice: A. Mouralidarane, et al.; Hepatology 58, 128 (2013), Abstract;
Quercetin reduces oxidative damage induced by paraquat via modulating expression of antioxidant genes in A549 cells: T. Zerin, et al.; J. Appl. Toxicol. 33, 1460 (2013), Abstract;
Rutin Suppresses Palmitic Acids-Triggered Inflammation in Macrophages and Blocks High Fat Diet-Induced Obesity and Fatty Liver in Mice: M. Gao, et al.; Pharm. Res. 30, 2940 (2013), Application(s): Total ROS determined in murine macrophages cell lines, Abstract;
Deoxycholic acid causes DNA damage while inducing apoptotic resistance through NF-{kappa}B activation in benign Barrett's epithelial cells: X. Huo, et al.; Am. J. Physiol. Gastrointest. Liver Physiol. 301, G278 (2011), Abstract;
Depletion of cytosolic or mitochondrial thioredoxin increases CYP2E1-induced oxidative stress via an ASK-1-JNK1 pathway in HepG2 cells: L. Wang, et al.; Free Radic. Biol. Med. 51, 185 (2011), Abstract;
Enhancement of the radiation effects by D-allose in head and neck cancer cells: H. Hoshikawa, et al.; Cancer Lett. 306, 60 (2011), Abstract;
Formation of TiO2 Nanostructures by Enzyme-Mediated Self-Assembly for the Destruction of Macrophages : K. Hayashi, et al.; Chem. Mater. 23, 3341 (2011), Abstract;
Protective effects of cynaroside against H2O2-induced apoptosis in H9c2 cardiomyoblasts: X. Sun, et al.; J. Cell. Biochem. 112, 2019 (2011), Abstract;
CYP2E1 enhances ethanol-induced lipid accumulation but impairs autophagy in HepG2 E47 cells: D. Wu, et al; Biochem. Biophys. Res. Commun. 402, 116 (2010), Abstract;

General Literature References

Fluorescent and luminescent probes for measurement of oxidative and nitrosative species in cells and tissues: progress, pitfalls, and prospects: P. Wardman; Free Radic. Biol. Med. 43, 995 (2007), Abstract;
Fluorescence probes used for detection of reactive oxygen species: A. Gomes, et al.; J. Biochem. Biophys. Methods 65, 45 (2005), Abstract;
Determination of mitochondrial reactive oxygen species: methodological aspects: C. Batandier, et al.; J. Cell. Mol. Med. 6, 175 (2002), Abstract;
Methods of detection of vascular reactive species: nitric oxide, superoxide, hydrogen peroxide, and peroxynitrite: M.M. Tarpey & I. Fridovich; Circ. Res. 89, 224 (2001), Abstract;

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