Directly monitors reactive oxygen and/or nitrogen species (ROS/RNS) in live cells
Discriminates among superoxide, nitric oxide and peroxynitrite
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/RNS inducers and scavengers
Enzo Life Sciences ROS-ID® ROS/RNS detection kit is designed to directly monitor real time reactive oxygen and/or nitrogen species (ROS/RNS) production in live cells using fluorescence microscopy. The kit includes three detection probes: NO Detection Reagent (Red, Ex/Em 650/670 nm), Oxidative Stress Detection Reagent (Green, Ex/Em 490/525 nm) for total ROS detection, and Superoxide Detection Reagent (Orange, Ex/Em 550/620 nm). It also includes the positive controls, pyocyanin and L-arginine, which are common inducers of ROS and NO production, respectively, as well as the negative controls, N-acetyl-L-cysteine and c-PTIO (common scavengers of ROS and NO, respectively). Through the combination of the three detection probes with a set of specific inhibitors and activators, this kit enables discrimination among superoxide, nitric oxide and peroxynitrite.
Excitation and emission spectra of Oxidative Stress Detection Reagent (green), Superoxide Detection Reagent (blue), and NO Detection Reagent (red).
Representative experiment demonstrating peroxynitrite and NO generation in HeLa cells using the ROS/RNS Detection Kit. The analysis demonstrates that combined treatment with L-arginine and pyocyanin generates peroxynitrite (green fluorescence), due to the reaction of NO with superoxide, but little NO (red fluorescence).
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Product Details
Alternative Name:
Reactive oxygen species / Reactive nitrogen species
Applications:
Fluorescence microscopy
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.
Quality Control:
ROS-ID® ROS/RNS 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), orange (550/620 nm), and red (650/670 nm) fluorescent cubes. Stained HeLa cells induced with L-arginine exhibit bright red fluorescence with a red punctuate cytoplasmic staining pattern. L-arginine-induced cells incubated with NO Scavenger (c-PTIO) demonstrate minimal red fluorescence comparable to control values. Stained HeLa cells induced with pyocyanin exhibit bright orange signal in the nucleus as well as bright green fluorescence in the cytoplasm, but no red signal specific for nitric oxide. Pyocyanin-induced cells incubated with ROS Inhibitor (N-acetyl-L-cysteine) demonstrate minimal green or orange fluorescence signal comparable to control values.
Quantity:
200 assays
Use/Stability:
With proper storage, the kit components are stable up to the date noted on the product label. Store the kit at -20°C in a non-frost free freezer, or -80°C for longer term storage.
Free radicals and other reactive species play seminal roles in many psychological and pathophysiological processes. Once produced within a cell, free radicals can damage a wide variety of cellular constituents, including proteins, lipids and DNA. However, at lower concentrations these very same agents may serve as second messengers in cellular signaling. Information-rich methods are required to quantify the relative levels of various reactive species.
Technical Info/Product Notes:
The ROS-ID® ROS/RNS 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.
Regulatory Status:
RUO - Research Use Only
Product Literature References
Hypoxia-induced oxidative stress promotes therapy resistance via upregulation of heme oxygenase-1 in multiple myeloma: K. Abe, et al.; Cancer Med. 12, 9709 (2023), Abstract;
Dihydroartemisinin-transferrin adducts enhance TRAIL-induced apoptosis in triple-negative breast cancer in a P53-independent and ROS-dependent manner: X. Zhou, et al.; Front. Oncol. 11, 789336 (2022), Abstract; Full Text
Intermittent exposure of cultured endothelial cells to physiologically relevant fructose concentrations has a profound impact on nitric oxide production and bioenergetics: M.L. Fiorello, et al.; PLoS One 17, e0267675 (2022), Abstract;
S1P (Sphingosine-1-Phosphate)-Induced Vasodilation in Human Resistance Arterioles During Health and Disease: B. Katunaric, et al.; Hypertension 79, 2250 (2022), Abstract;
Extra-mitochondrial citrate synthase initiates calcium oscillation and suppresses age-dependent sperm dysfunction: W. Kang, et al.; Lab. Invest. 100, 583 (2020), Abstract; Full Text
Liposome-encapsulated bacillus calmette-guérin cell wall skeleton enhances antitumor efficiency for bladder cancer in vitro and in vivo via induction of amp-activated protein kinase: Y.M. Whang, et al.; Cancers (Basel) 12, 3679 (2020), Abstract; Full Text
Mesenchymal stem cell conditioned medium ameliorates diabetic serum-induced insulin resistance in 3T3-L1 cells: A. Sanap, et al.; Chronic Dis. Transl. Med. 7, 47 (2020), Abstract; Full Text
Isoorientin derived from Gentiana veitchiorum Hemsl. flowers inhibits melanogenesis by down-regulating MITF-induced tyrosinase expression: Q. Wu, et al.; Phytomedicine 57, 129 (2019), Application(s): Confocal microscopy; mouse melanoma cell line (B16-F10), Abstract;
Low intensity ultrasound reduces high glucose reduced nitric oxide generation in retinal pigment epithelial cells: M.B. Karmacharya, et al.; Ultrasound Med. Biol. 44, 647 (2018), Application(s): Fluorescence microscopy, Abstract;
HEK-293 secretome attenuates kainic acid neurotoxicity through insulin like growth factor-phosphatidylinositol-3-kinases pathway and by temporal regulation of antioxidant defense machineries: C. Venugopal, et al.; Neurotoxicology 69, 189 (2017), Application(s): ROS detection in Murine Hippocampal cells, Abstract;
Selective enhancing effect of metal ions on mutagenicity: N. Fujii, et al.; Genes Environ. 38, 21 (2016), Application(s): Intracellular reactive oxygen species, Abstract; Full Text
Cell-to-cell diffusion of glucose in the mammalian heart is disrupted by high glucose. Implications for the diabetic heart: W.C. De Mello ; Exp. Cell Res. 334, 239 (2015), Application(s): Fluorescence Microscopy, Abstract;
Increased Proinflammatory Cytokine Production and Decreased Cholesterol Efflux Due to Downregulation of ABCG1 in Macrophages Exposed to Indoxyl Sulfate: K. Matsuo, et al. ; Toxins (Basel) 7, 3155 (2015), Application(s): Measurement of reactive oxygen species production, Abstract; Full Text
Regulation of gene expression by tobacco product preparations in cultured human dermal fibroblasts: G.E. Malpass, et al.; Toxicol. Appl. Pharmacol. 279, 211 (2014), Application(s): Microplate measurements of ROS and RNS levels in human dermal fibroblasts, Abstract;
Enterococcus faecalis infection causes inflammation, intracellular oxphos-independent ROS production, and DNA damage in human gastric cancer cells: J.A. Strickertsson, et al.; PLoS One 8, e63147 (2013), Application(s): Observation of ROS/RNS production by confocal microscopy, Abstract; Full Text
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;
