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ROS-ID® ROS/RNS detection kit

Monitor  reactive oxygen species and nitrogen species in live cells
 
ENZ-51001-200 1 Kit 281.00 USD
Do you need bulk/larger quantities?
 
  • 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 fluorescent dye reagents: NO Detection Reagent (red fluorescent), Oxidative Stress Detection Reagent (green fluorescent) for total ROS detection, and Superoxide Detection Reagent (orange fluorescent). 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.
ENZ-51001 HELA
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).

ENZ-51001 Spectra OSDa
Excitation and emission spectra of Oxidative Stress Detection Reagent (green), Superoxide Detection Reagent (blue), and NO Detection Reagent (red).
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ENZ-51001 HELA ENZ-51001 Spectra OSDa

Product Specification

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.
 
Handling:Protect from light. Avoid freeze/thaw cycles.
 
Shipping:Shipped on Dry Ice
 
Long Term Storage:-80°C
 
Kit/Set Contains:NO Detection Reagent (Red), 60µl
Oxidative Stress Detection Reagent (Green), 300 nmoles
Superoxide Detection Reagent (Orange), 300 nmoles
NO Inducer (L-Arginine), 100 µl
ROS Inducer (Pyocyanin), 1 µmole
NO Scavenger (c-PTIO), 400 nmoles 
ROS Inhibitor (N-acetyl-L-cysteine), 2x10 mg
10X Wash Buffer, 15 ml
 
Scientific Background: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.
 

Product Literature References

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;

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