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.
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.
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.
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.
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;
