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ROS-ID® Total ROS detection kit

ENZ-51011 1 Kit 175.00 USD
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  • Directly monitors global levels of reactive oxygen species (ROS), but not superoxide in live cells
  • Distinguishes between different reactive species, such as hydrogen peroxide, peroxynitrite and hydroxyl radicals when specific inhibitors are used
  • 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
  • Stringently manufactured, to control and eliminate non-specific assay artifacts
Enzo Life Sciences’ ROS-ID® Total ROS detection 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. The kit includes Oxidative Stress Detection Reagent (Green) as the major component. This non-fluorescent, cell-permeable total ROS detection dye reacts directly with a wide range of reactive species, such as hydrogen peroxide, peroxynitrite and hydroxyl radicals, yielding a green fluorescent product indicative of cellular production of different ROS/RNS types. The kit is not designed to detect superoxide, reactive chlorine or bromine species, as the fluorescent probe included is relatively insensitive to these analytes. 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. The Total ROS Detection Kit contains sufficient reagents for at least 200 microscopy assays or 50 flow cytometry assays using live cells (adherent or in solution).
ENZ-51011 Fig1
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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ENZ-51011 Fig1

Product Specification

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:Shipped on 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

Product Literature References

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
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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Review for Total ROS detection kit for microscopy and flow cytometry

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