Online Purchasing Account You are logged on as Guest. LoginRegister a New AccountShopping cart (Empty)
United States 

ROS-ID® Total ROS/Superoxide detection kit

Directly Monitor Global Levels  of Reactive Oxygen Species (ROS), and Specifically Superoxide
 
ENZ-51010 1 Kit 235.00 USD
Do you need bulk/larger quantities?
 
  • Distinguishes between different reactive species, such as hydrogen peroxide, peroxynitrite and hydroxyl radicals
  • High sensitivity, specificity and accuracy for live cell studies
  • Compatible with major components of tissue culture media (phenol red, FBS and BSA)
  • Complete set of reagents, including ROS inducers and scavengers
  • Validated on microscopy, flow cytometry, and microplates
Enzo Life Sciences’  ROS-ID® Total ROS/Superoxide detection kit is designed to directly monitor real time reactive oxygen species (ROS) production in live cells using fluorescence microscopy or flow cytometry. The kit includes two fluorescent dyes as major components: Oxidative Stress Detection Reagent (Green) for total ROS detection reagent and Superoxide Detection Reagent (Orange). Through the combination of these two specific fluorescent probes, the kit provides a simple and specific assay for the real-time measurement of global levels of reactive oxygen species (ROS), and specifically superoxide in living cells. The Total ROS/Superoxide Detection Kit contains sufficient reagents for at least 200 microscopy assays or 50 flow cytometry assays using live cells (adherent or in suspension) or 2x96 microplate assays.
ENZ-51010 Fig1
Figure 1. Profiling of reactive oxygen species formation by fluorescence microscopy was achieved in HeLa cells loaded with ROS/Superoxide detection reagents and treated with pyocyanin. General oxidative stress levels were monitored in the green channel, while superoxide production was detected in the orange channel. Pretreatment with NAC, a general ROS inhibitor, prevents formation of ROS.
ENZ-51010 FC
Figure 2. Jurkat cells were induced with 100 µM pyocyanin (general ROS inducer, panel B), 200 µM antimycin A (superoxide inducer, panel C) or 1 µM of t-butyl-hydroperoxide (peroxide inducer, panel D), stained with two color ROS Detection Kit and analyzed using flow cytometry. Untreated cells (panel A) were used as a control. Cell debris were ungated and compensation was performed using single stained pyocyanin-treated samples. Red numbers reflect the percentage of the cells in each quadrant.
ENZ-51010 NAC
Figure 3. Flow cytometry histograms of ROS inhibition by N-acetyl-L-cysteine (NAC, ROS inhibitor). Jurkat cells were pretreated with 5 mM (pink, left) or 10 mM (pink, right), or without NAC (green, positive control) for one hour. Cells were then incubated with 100 µM pyocyanin (ROS inducer) and stained with 1 µM oxidative stress detection reagent. Untreated cells (blue) were used as a negative control. The numbers in the upper right corners indicate the percentage of median fluorescence intensity of NAC-treated cells as compared with positive control cells.
ENZ-51010 bar graph
Figure 4. Profiling of ROS formation by flow cytometry in Hela cells. Data represents % positive following treatment with Pyocyanin (ROS/SO inducer), TBHP (ROS inducer), and AMA (superoxide inducer). Green columns indicate % positive for ROS and orange columns represent % positive for superoxide.
ENZ-51010 double-chart
Figure 5. (Top) HeLa Cells Loaded with ROS/RNS 3-Plex Detection Reagent for 2 h, 37°C and Induced for 20 min, 37°C.
Column 1: Control
Column 2: Pyocyanin, 0.1 mM
Column 3: L-Arginine, 1mM
Column 4: Pyo/Arg

(Bottom) Pretreatment with 5 mM NAC
Column 1: Control
Column 2: L-Arginine, 1mM
Column 3: Pyocyanin, 0.1 mM
Column 4: Pyo/Arg

Specific profiling of reactive oxygen/nitrogen species formation by fluorescence microscopy in HeLa cells loaded with dyes from ROS-ID® ROS/RNS Detection Kit (ENZ-51001). (Top) - Combined treatment of Pyocyanin and L-Arg generates peroxynitrite (green fluorescence) due to the reaction of NO with superoxide, bur little NO (red fluorescence). (Bottom) - Pretreatment with NAC inhibits peroxynitrite and superoxide formation, but not NO.
ENZ-51010 doublegraphs
Figure 6. ROS-ID® Total ROS/Superoxide detection Kit and a set of ROS scavengers/inhibitors were used to profile ROS production in HeLa cells treated with antimycin A, (AMA, specific superoxide inducer), t-butyl-peroxide (TBHP, specific peroxide inducer), and pyocyanin (general ROS inducer). Similar results were obtained using U-2 OS and CHO K1 cells.
ENZ-51010 Kit Box
Please mouse over
ENZ-51010 Fig1 ENZ-51010 FC ENZ-51010 NAC ENZ-51010 bar graph ENZ-51010 double-chart ENZ-51010 doublegraphs ENZ-51010 Kit Box

Product Specification

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/Superoxide 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 (490/525 nm) and orange (550/620 nm) filter set.
The following results are obtained: ROS positive control samples induced with Pyocyanin exhibit bright orange signal in nucleus as well as bright green fluorescence in the cytoplasm. Cells pretreated with the ROS inhibitor don’t demonstrate any green or orange fluorescence signal upon induction.
 
Quantity:200 fluorescence microscopy assays or 50 flow cytometry assays or 2x96 microplate 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:Shipped on Dry Ice
 
Short Term Storage:-20°C
 
Long Term Storage:-80°C
 
Kit/Set Contains:Oxidative Stress Detection Reagent (Green), 300 nmoles
Superoxide Detection Reagent (Orange), 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/Superoxide 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. CELLELSTIAL® 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.

Toxicology Application Note:
Use of 3D Cultured Human iPSC-Derived Hepatocytes for Long-Term Hepatotoxicity Studies
Screening Reactive Oxygen Species (ROS) on IQUE® Screener

Cited samples:
For an overview on cited samples please click here.
 

Product Literature References

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;
Neurochemical effects of the R form of α-lipoic acid and its neuroprotective mechanism in cellular models of Parkinson's disease: H. Zhao, et al.; Int. J. Biochem. Cell. Biol. 87, 86 (2017), Application(s): Flow cytometry using SH-SY5Y cells, Abstract;
Polyethylene glycol-functionalized poly (Lactic Acid-co-Glycolic Acid) and graphene oxide nanoparticles induce pro-inflammatory and apoptotic responses in Candida albicans-infected vaginal epithelial cells: R.D. Wagner, et al.; PLoS One 12, e0175250 (2017), Abstract; Full Text
Isovitexin Exerts Anti-Inflammatory and Anti-Oxidant Activities on Lipopolysaccharide-Induced Acute Lung Injury by Inhibiting MAPK and NF-κB and Activating HO-1/Nrf2 Pathways: H. Lv, et al.; Int. J. Biol. Sci. 12, 72 (2016), Application(s): measured Intracellular ROS lebels by Fluorence microplate reader or flow cytometry, Abstract; Full Text
Nilotinib induces ER stress and cell death in H9c2 cells: D. Lekes, et al.; Physiol. Res. 65, S505 (2016), Application(s): Fluorescence microscopy using H9c2 cells, Abstract; Full Text
Prolactin protects retinal pigment epithelium by inhibiting sirtuin 2-dependent cell death: R. Meléndez García, et al.; EBioMedicine 7, 35 (2016), Application(s): Measurement of total ROS, Abstract;
Salubrinal reduces oxidative stress, neuroinflammation and impulsive-like behavior in a rodent model of traumatic brain injury: A.F. Logsdon, et al.; Brain Res. 1643, 140 (2016), Application(s): Detected total ROS and superoxide levels in rodent cells, Abstract;
Acinetobacter baumannii escape from neutrophil extracellular traps (NETs): G. Kamoshida, et al.; J. Infect. Chemother. 21, 43 (2015), Application(s): Flow cytometry measurements of total ROS and superoxide levels in neutrophils, Abstract;
Anthocyans-rich Aronia melanocarpa extract possesses ability to protect endothelial progenitor cells against angiotensin II induced dysfunction: A. Parzonko, et al.; Phytomedicine 22, 1238 (2015), Application(s): Measurement of intracellular ROS production, Abstract;
Cytotoxicity of the Urokinase-Plasminogen Activator Inhibitor Carbamimidothioic Acid (4-Boronophenyl) Methyl Ester Hydrobromide (BC-11) on Triple-Negative MDA-MB231 Breast Cancer Cells: A. Longo, et al.; Molecules 20, 9879 (2015), Application(s): Flow Cytometry, Abstract; Full Text
Functional pseudogenes inhibit the superoxide production: W. Xu, et al.; Precis. Med. 2, e745 (2015), Application(s): Fluorescence Microscopy, Abstract; Full Text
Gemcitabine-induced CXCL8 expression counteracts its actions by inducing tumor neovascularization: Y. Song, et al.; Biochem. Biophys. Res. Commun. 458, 341 (2015), Application(s): Flow Cytometry, Abstract;
Mechanisms that lead to the regulation of NLRP3 inflammasome expression and activation in human dental pulp fibroblasts: A. Zhang, et al.; Mol. Immunol. 66, 253 (2015), Application(s): ELISA using human cells, Abstract;
Mitofusin-2 triggers mitochondria Ca 2+ influx from the endoplasmic reticulum to induce apoptosis in hepatocellular carcinoma cells: W. Wang, et al.; Cancer Lett. 358, 47 (2015), Application(s): Flow cytometry measurements of total ROS and superoxide levels in HepG2 cells, Abstract;
Neither resveratrol nor metformin protects human cardiomyocytes against toxicity of epirubicin and radiation in-vitro: M. Gramatyka, et al.; Med. Sci. 17, 1 (2015), Application(s): Microplate, Full Text
Oxidative Stress Response in Canine In Vitro Liver, Kidney and Intestinal Models with Seven Potential Dietary Ingredients: K. Choi, et al. ; Toxicol. Lett. 241, 49 (2015), Application(s): Oxidative/nitrosative stress measurements for hepatocytes, Abstract;
Phenotypic screening of a library of compounds against metastatic and non-metastatic clones of a canine mammary gland tumour cell line: K. Saeki, et al.; Vet. J. 205, 288 (2015), Abstract;
A feedback mechanism to control apoptosis occurs in the digestive gland of the oyster crassostrea gigas exposed to the paralytic shellfish toxins producer Alexandrium catenella: J.L. Rolland, et al.; Mar. Drugs 12, 5035 (2014), Application(s): Microplate measurements of total ROS and superoxide levels in digestive gland cells from oysters, Abstract; Full Text
A new and reliable method for live imaging and quantification of reactive oxygen species in Botrytis cinerea: technological advancement: R. Marschall, et al.; Fungal Genet. Biol. 71, 68 (2014), Application(s): Fluorescence microscopy and microplate measurements of total ROS and superoxide levels in fungi , Abstract;
Dengue virus type 2 (DENV2)-induced oxidative responses in monocytes from glucose-6-phosphate dehydrogenase (G6PD)-deficient and G6PD normal subjects: A.A. Al-Alimi, et al.; PloS Negl. Trop. Dis. 8, e2711 (2014), Application(s): Flow cytometry measurements of total ROS and superoxide levels in monocytes, Abstract; Full Text
Ghrelin protects against renal damages induced by angiotensin-II via an antioxidative stress mechanism in mice: K. Fujimura, et al.; PLoS One 9, e94373 (2014), Application(s): Microplate measurements of total ROS and superoxide levels in HK-2 cells, Abstract; Full Text
Metabolic reprogramming of macrophages: glucose transporter 1 (GLUT1)-mediated glucose metabolism drives a proinflammatory phenotype: A.J. Freemerman, et al.; J. Biol. Chem. 289, 7884 (2014), Application(s): Flow cytometry measurements of total ROS and superoxide levels in macrophages, Abstract;
Mice with heterozygous deficiency of manganese superoxide dismutase (SOD2) have a skin immune system with features of "inflamm-aging": J. Scheurmann, et al.; Arch. Dermatol. Res. 306, 143 (2014), Abstract;
Nuclear Nrf2 induction by protein transduction attenuates osteoclastogenesis: H. Kanzaki, et al.; Free Radic. Biol. Med. 77, 239 (2014), Application(s): Flow cytometry measurements of total ROS and superoxide levels in macrophages, Abstract;
Rhododendrol, a depigmentation-inducing phenolic compound, exerts melanocyte cytotoxicity via a tyrosinase-dependent mechanism: M. Sasaki, et al.; Pigment Cell Melanoma Res. 27, 754 (2014), Application(s): Flow cytometry measurements of total ROS and superoxide levels in melanocytes, Abstract; Full Text
The medicinal fungus Antrodia cinnamomea suppresses inflammation by inhibiting the NLRP3 inflammasome: T.T. Huang, et al.; J. Ethnopharmacol. 155, 154 (2014), Application(s): Microplate measurements of total ROS and superoxide levels in THP-1 macrophages, Abstract;
Translocation of GSK-3β, a trigger of permeability transition, is kinase activity-dependent and mediated by interaction with VDAC2: M. Tanno, et al.; J. Biol. Chem. 289, 29285 (2014), Application(s): Fluorescence microscopy of total ROS and superoxide production in H9c2 and HEK293 cells, Abstract;
Ubiquinol-10 supplementation activates mitochondria functions to decelerate senescence in senescence-accelerated mice: G. Tian, et al.; Antioxid. Redox Signal. 20, 2606 (2014), Abstract;
Validation of genome-wide association study (GWAS)-identified disease risk alleles with patient-specific stem cell lines: J. Yang, et al.; Hum. Mol. Genet. 23, 3445 (2014), Abstract;
Vasoinhibins regulate the inner and outer blood-retinal barrier and limit retinal oxidative stress: D. Arredondo-Zamarripa, et al.; Front. Cell. Neurosci. 8, 333 (2014), Application(s): Microplate measurements of total ROS and superoxide levels in bovine umbilical vein endothelial cells and ARPE-19 cells, Abstract; Full Text
Connexin36 contributes to INS-1E cells survival through modulation of cytokine-induced oxidative stress, ER stress and AMPK activity: F. Allagnat, et al.; Cell Death Differ. 20, 1742 (2013), Application(s): Measurement of ROS and Superoxide using a microplate reader, Abstract;
Dual regulation of cadmium-induced apoptosis by mTORC1 through selective induction of IRE1 branches in unfolded protein response: H. Kato, et al.; PLoS One 8, e64344 (2013), Application(s): Fluorescence microscopy using a rat renal tubular epithelial cell line, Abstract; Full Text
Embryonic stem cell-based modeling of tau pathology in human neurons: J. Mertens, et al.; Am. J. Pathol. 182, 1769 (2013), Application(s): Measurement of total ROS/Superoxides in human neurons using microplate reader, Abstract;
Growth inhibition and apoptosis of neuroblastoma cells through ROS-independent MEK/ERK activation by sulforaphane: Y.C. Hsu, et al.; Cell. Biochem. Biophys. 66, 765 (2013), Abstract;
Metabolomic analysis of dynamic response and drug resistance of gastriccancer cells to 5-fluorouracil: S. Sasada, et al.; Oncol. Rep. 29, 925 (2013), Application(s): Flow cytometry using a gastric adenocarcinoma cell line line, Abstract; Full Text
Mitochondrial dysfunction in a novel form of autosomal recessive ataxia: N.A. Murad, et al.; Mitochondrion 13, 235 (2013), Application(s): Flow cytometry using fibroblasts, Abstract;
Oleuropein and oleacein may restore biological functions of endothelial progenitor cells impaired by angiotensin II via activation of Nrf2/heme oxygenase-1 pathway: A. Parzonko, et al.; Phytomedicine 20, 1088 (2013), Application(s): Measurement of total ROS/Superoxides in human PBMC using microplate reader, Abstract;
Reduction of connexin36 content by ICER-1 contributes to insulin-secreting cells apoptosis induced by oxidized LDL particles: J.A. Haefliger, et al.; PLoS One 8, e55198 (2013), Application(s): Measurement of total ROS/Superoxides in a rat insulinoma cell line using microplate reader, Abstract; Full Text
The Keap1/Nrf2 axis plays a role in osteoclast differentiation by regulating intracellular ROS signaling: H. Kanzaki, et al.; J. Biol. Chem. 288, 23009 (2013), Application(s): Flow cytometry using mouse cells, Abstract; Full Text
Aberrant methylation of PSD disturbs Rac1-mediated immune responses governing neutrophil chemotaxis and apoptosis in ulcerative colitis-associated carcinogenesis: T. Kato, et al.; Int. J. Oncol. 40, 942 (2012), Application(s): Fluorescence microscopy using a human skin fibroblast cell line, Abstract; Full Text
Cytotoxic effects of Jay Amin hydroxamic acid (JAHA), a ferrocene-based class I histone deacetylase inhibitor, on triple-negative MDA-MB231 breast cancer cells: M. Librizzi, et al.; Chem. Res. Toxicol. 25, 2608 (2012), Abstract;
Global DNA methylation levels are modulated by mitochondrial DNA variants: D. Bellizzi, et al.; Epigenomics 4, 17 (2012), Abstract;
Manufacturing strategy for multiwalled carbon nanotubes as a biocompatible and innovative material: H. Haniu, et al.; Journal of Nanotechnology 2012, 1 (2012), Application(s): FC using human bronchial epithelial cells, Full Text
Metabolic remodeling precedes mitochondrial outer membrane permeabilization in human glioma xenograft cells: S. Ponnala, et al.; Int. J. Oncol. 40, 509 (2012), Application(s): Flow cytometry and fluorescence microscopy using human glioma xenograft cells, Abstract; Full Text
Phenylalkyl isoselenocyanates vs phenylalkyl isothiocyanates: thiol reactivity and its implications: M.A. Crampsie, et al.; Chem. Biol. Interact. 200, 28 (2012), Application(s): Flow cytometry using A549 cells, Abstract;
Glucose and collagen regulate human platelet activity through aldose reductase induction of thromboxane: W.H. Tang, et al.; J. Clin. Invest. 121, 4462 (2011), Abstract; Full Text
Activation of the p38 MAPK/Akt/ERK1/2 signal pathways is required for the protein stabilization of CDC6 and cyclin D1 in low-dose arsenite-induced cell proliferation: Y. Liu, et al.; J. Cell. Biochem. 111, 1546 (2010), Abstract;

Related Products

ROS-ID® Total ROS detection kit 

Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS | Print as PDF
 
ENZ-51011 1 Kit 172.00 USD
Do you need bulk/larger quantities?
 

ROS-ID® Superoxide detection kit 

Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS | Print as PDF
 
ENZ-51012 1 Kit 172.00 USD
Do you need bulk/larger quantities?
 

Red hydrogen peroxide assay kit  

Fluorescence microscopy, HTS | Print as PDF
 
ENZ-51004 1 Kit 289.00 USD
Do you need bulk/larger quantities?
 

ROS-ID® ROS/RNS detection kit 

Monitor  reactive oxygen species and nitrogen species in live cells
Fluorescence microscopy | Print as PDF
 
ENZ-51001-200 1 Kit 281.00 USD
Do you need bulk/larger quantities?
 

Luminol (ultra pure) 

ROS dye
521-31-3, ≥95% (HPLC) | Print as PDF
 
ENZ-52354 1 g 96.00 USD
Do you need bulk/larger quantities?
 

ROS-ID® Hypoxia/Oxidative stress detection kit 

Specific detection system for simultaneous analysis of hypoxia and oxidative stress using microscopy and flow cytometry
Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS | Print as PDF
 
ENZ-51042-0125 125 tests 105.00 USD
 
ENZ-51042-K500 500 tests 367.00 USD
Do you need bulk/larger quantities?
 

Related Literature

Brochures
Comprehensive Tools for Quantifying Cellular Responses to Oxidative Damage
Comprehensive Tools for Quantifying Cellular Responses to Oxidative Damage
Download as PDF

Brochures
Predictive Toxicology
Predictive Toxicology
Download as PDF

Scientific Posters
Kinetic Profiling of Reactive Oxygen Species Generation in Living Cells Using A Simple and Robust Fluorescence Microplate-based Assay Workflow
Kinetic Profiling of Reactive Oxygen Species Generation in Living Cells Using A Simple and Robust Fluorescence Microplate-based Assay Workflow
Download as PDF

Product Flyers
ROS-ID® Total ROS/Superoxide detection kit
ROS-ID® Total ROS/Superoxide detection kit
Download as PDF

Product Flyers
New Tools for Flow Cytometry
New Tools for Flow Cytometry
Download as PDF

All new literature pieces

Recommend this page

 
For Research Use Only. Not for use in diagnostic procedures.
Keep in touch

©2017 Enzo Life Sciences, Inc.,