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ROS-ID® NO Detection kit

ENZ-51013-200 1 Kit 204.00 USD
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  • Directly monitor real time NO production in live cells by fluorescence microscopy
  • High specificity, sensitivity and accuracy, yielding a water-insoluble red fluorescent product
  • Distinguish between peroxynitrite and nitric oxide
  • NO inducer and scavenger included
Enzo Life Sciences’ ROS-ID® NO Detection kit is designed to directly monitor real time NO production in live cells by fluorescence microscopy using NO Detection Reagent (red fluorescent) as the major component. The non-fluorescent, cell-permeable NO detection dye reacts with NO in the presence of O2 with high specificity, sensitivity and accuracy, yielding a water-insoluble red fluorescent product. Importantly, this dye is not reactive towards peroxynitrite that allows distinguishing between peroxynitrite and nitric oxide. The kit is not designed to detect reactive chlorine or bromine species, as the fluorescent probes included are relatively insensitive to these analytes. Upon staining, the fluorescent products generated by this dye can be visualized using a wide-field fluorescence microscope equipped with any red fluorescent cube. The combination of 650/670 nm is recommended when additional fluorescence signals (green or orange) would be detected simultaneously The NO Detection Kit contains sufficient reagents for at least 200 microscopy assays using live cells (adherent or in suspension).

Product Details

Applications:Fluorescence microscopy, Fluorescent detection
Application Notes:This kit is designed to directly monitor real time NO production in live cells using fluorescence microscopy.
Quality Control:A sample from each lot of ROS-ID® NO 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 red (650/670 nm) fluorescent cubes.
The following results are obtained: Nitric Oxide positive control samples induced with L-Arginine exhibit red fluorescence with a red punctuate cytoplasmic staining pattern. Cells incubated with the NO Scavenger (c-PTIO), post induced with L-Arginine don’t demonstrate any red fluorescence.
Quantity:200 fluorescence microscopy assays.
Handling:Protect from light. Avoid freeze/thaw cycles.
Shipping:Shipped on Dry Ice
Short Term Storage:-20°C
Long Term Storage:-80°C
Contents:NO Detection Reagent (Red), 60 µl
NO Inducer (L-Arginine), 100 µl
NO Scavenger (c-PTIO), 400 nmoles
10X Wash Buffer, 15 ml 
Technical Info/Product Notes:The ROS-ID® NO 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

Human M1 macrophages express unique innate immune response genes after mycobacterial infection to defend against tuberculosis: A. Karshan, et al.; Commun. Biol. 5, 480 (2022), Abstract;
High-content analysis monitoring intracellular trafficking and replication of Mycobacterium tuberculosis inside host cells: N. Debousere, et al.; Methods Mol. Biol. 2314, 649 (2021), Abstract;
Hyperhomocysteinemia and Low Folate and Vitamin B12 Are Associated with Vascular Dysfunction and Impaired Nitric Oxide Sensitivity in Morbidly Obese Patients: M. Haloul, et al.; Nutrients 12, 2014 (2020), Abstract; Full Text
Stimulation of TRPA1 attenuates ischemia-induced cardiomyocyte cell death through an eNOS-mediated mechanism: S.R. Andrei, et al.; Channels (Austin) 13, 192 (2019), Abstract; Full Text
Anti-inflammatory effects of olive-derived hydroxytyrosol on lipopolysaccharide-induced inflammation in RAW264.7 cells: Y. Yonezawa, et al.; J. Vet. Med. Sci. 80, 1801 (2018), Abstract; Full Text
Oligodendrocyte RasG12V expressed in its endogenous locus disrupts myelin structure through increased MAPK, nitric oxide, and notch signaling: H.E. Titus, et al.; Glia 65, 1990 (2017), Application(s): Flow cytometry with murine brain cells., Abstract; Full Text
Improved arterial flow-mediated dilation after exertion involves hydrogen peroxide in overweight and obese adults following aerobic exercise training: A.T. Robinson, et al.; J. Hypertens. 34, 1309 (2016), Application(s): Microscopy on human microvessels, Abstract;
STAT3 Represses Nitric Oxide Synthesis in Human Macrophages upon Mycobacterium tuberculosis Infection: C.J. Queval, et al.; Sci. Rep. 6, 29297 (2016), Abstract; Full Text
Flavonoids activate endothelial nitric oxide synthase by altering their phosphorylation via mitogen-activated protein kinase pathways in glucose-induced endothelial cells: C.E. Kim, et al.; J. Funct. Foods 17, 676 (2015), Application(s): HUVECs,
Ginsenoside Rg3 regulates S-Nitrosylation of the NLRP3 inflammasome via suppression of iNOS: S. J. Yoon, et al.; Biochem. Biophys. Res. Commun. 463, 1184 (2015), Application(s): ELISA using mouse serum, liver and spleen samples, Abstract;
Propofol restores TRPV1 sensitivity via a TRPA1-, nitric oxide synthase-dependent activation of PKCε: P. Sinharoy, et al.; Pharmacol. Res. Perspect. 3, e00153 (2015), Application(s): Fluorescence microscopy to detect NO in F-11 cells, Abstract; Full Text
Reduced flow-and acetylcholine-induced dilations in visceral compared to subcutaneous adipose arterioles in human morbid obesity: I. Grizelj, et al.; Microcirculation 22, 44 (2015), Abstract;
Anti-inflammatory effects of Edaravone and Scutellarin in activated microglia in experimentally induced ischemia injury in rats and in BV-2 microglia: Y. Yuan, et al.; BMC Neurosci. 15, 125 (2014), Application(s): Fluorescence microscopy to detect NO in BV-2 microglia cells, Abstract; Full Text
Lipopolysaccharide (LPS) stimulation of fungal secondary metabolism: Z.G. Khalil, et al.; Mycology 5, 168 (2014), Application(s): Fluorescence microscopy to detect NO in fungal cells, Abstract; Full Text
Cyclosporine attenuates arginine transport, in human endothelial cells, through modulation of cationic amino acid transporter-1: A. Grupper, et al.; Am. J. Nephrol. 37, 613 (2013), Abstract;
Identification and characterization of a functional mitochondrial angiotensin system: P.M. Abadir, et al.; PNAS 108, 14849 (2011), Application(s): NO measured in isolated mitochondria using microplate reader , Abstract;
Loss of bcl-2 during the senescence exacerbates the impaired angiogenic functions in endothelial cells by deteriorating the mitochondrial redox state: M. Uraoka, et al.; Hypertension 58, 254 (2011), Application(s): NO detected in HUVEC cells, 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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