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ROS-ID® Hypoxia/Oxidative stress detection kit

Specific detection system for simultaneous analysis of hypoxia and oxidative stress using microscopy and flow cytometry
 
ENZ-51042-0125 125 tests 105.00 USD
 
ENZ-51042-K500 500 tests 367.00 USD
Do you need bulk/larger quantities?
 
  • Specifically distinguishes hypoxia from oxidative stress in Real-time
  • High sensitivity, specificity and accuracy for live cell studies
  • Use with adherent or suspension cell lines
  • Complete set of reagents, including ROS and Hypoxia inducer controls.
Enzo Life Sciences’ ROS-ID® Hypoxia/Oxidative stress detection kit is designed for functional detection of hypoxia and oxidative stress levels in live cells (both suspension and adherent) using fluorescent microscopy or flow cytometry. This kit includes fluorogenic probes for hypoxia (red) and for oxidative stress levels (green) as two major components.

The Hypoxia (Red) dye takes advantage of the nitroreductase activity present in hypoxic cells by converting the Nitro group to hydroxylamine (NHOH) and amino (NH2) and releasing the fluorescent probe.

The Oxidative Stress Detection Reagent is a non-fluorescent, cell-permeable total ROS detection dye which reacts directly with a wide range of reactive species. The generated fluorescent products can be visualized using a wide-field fluorescence microscope equipped with standard fluorescein (490/525 nm) and Texas Red (596/670 nm) filters, confocal microscopy, or cytometrically using any flow cytometer equipped with a blue (488 nm) laser.
ENZ-51042 Fig1
The absorption and emission peaks for the Oxidative Stress (A) and Hypoxia Red (B) detection dyes are 504nm/524nm and 580nm/595nm, respectively. The dyes can be excited with an argon ion laser at 488 nm, and detected in the FL1 channel (Oxidative Stress dye) and FL3 Channel (Hypoxia Red dye) on ost bench flow cytometers.
ENZ-51042 Fig2
HeLa cells were subject to treatment. Bright red fluorescence of the Hypoxia probe is observed following its conversion by cellular nitroreductases under hypoxic conditions such as those induced chemically by treatment with the hypoxia-mimetic desferrioxamine (DFO).
ENZ-51042 Fig3
Detection of hypoxia and oxidative stress levels in cultured human HeLa and HL-60 cells. Cells were treated with hypoxia inducer (DFO) and ROS inducer (pyocyanin). Numbers in each quadrant reflects the percentage of cells (population). Results indicate that hypoxia and oxidative stress dye are specific
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ENZ-51042 Fig1 ENZ-51042 Fig2 ENZ-51042 Fig3

Product Specification

Applications:Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS
 
Application Notes:This kit is designed for fluorescence microscopy and/or flow cytometry using adherent or suspension cells.
 
Quality Control:The testing is accomplished using flow cytometry method for assessment of hypoxic cells and/or cells with high levels of total oxidative stress in conjunction with dyes (provided in kit). Microscopy images are also obtained.
 
Quantity:For -K500 size:
500 fluorescence microscopy assays or 100 flow cytometry assays.

For -0125 size:
125 fluorescence microscopy assays or 25 flow cytometry assays.
 
Use/Stability:With proper storage, the kit components are stable up to the date noted on the product label.
 
Handling:Avoid freeze/thaw cycles.
 
Shipping:Shipped on Dry Ice
 
Short Term Storage:-20°C
 
Long Term Storage:-20°C
 
Kit/Set Contains:Hypoxia Red Detection Reagent
Oxidative Stress Detection Reagent (Green)
ROS Inducer (Pyocyanin)
Hypoxia Inducer (DFO)
 
Technical Info/Product Notes:The ROS-ID® Hypoxia/Oxidative stress 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 cell analysis applications involving confocal microscopy, flow cytometry, microplate readers and HCS/HTS, where consistency and reproducibility are required.

Application Notes:
3D Culture Application: Inhibition of Hypoxic Tumor Cells using a Three-Dimensional Spheroid Model

3D Culture Application: PMT and Image-Based Analysis of Hypoxia Induction using a 3D Spheroid Model
 
Protocol:Detailed instructions are included in the manual for microscopy and flow cytometry applications for adherent and suspension cells.
 

Product Literature References

Investigating the application of a nitroreductase-expressing transgenic zebrafish line for high-throughput toxicity testing: A.C. Chlebowski, et al.; Toxicol. Rep. 4, 202 (2017), Application(s): Use of hypoxia detection reagent with zebrafish embryos,
Tumor-penetrating nanoparticles for enhanced anticancer activity of combined photodynamic and hypoxia-activated therapy: Y. Wang, et al.; ACS Nano 11, 2227 (2017), Application(s): Flow cytometry analysis of mouse breast carcinoma cells, Abstract; Full Text
Analysis of a nitroreductase-based hypoxia sensor in primary neuronal cultures: B.N. Lizama-Manibusan, et al.; ACS Chem. Neurosci. 7, 1188 (2016), Abstract;
ERK2 and CHOP restrict the expression of the growth-arrest specific p20K lipocalin gene to G0: M.J. Erb, et al.; Mol. Cell. Biol. 36, 2890 (2016), Application(s): Hypoxia levels in chick embryo fibroblasts (CEF), Abstract;
Phagocyte respiratory burst activates macrophage erythropoietin signalling to promote acute inflammation resolution: B. Luo, et al.; Nat. Commun. 7, 12177 (2016), Application(s): Flow cytometry analysis of hypoxia in exudate leukocytes and peritoneum, Abstract; Full Text
Selective advantage of trisomic human cells cultured in non-standard conditions: S.D. Rutledge, et al.; Sci. Rep. 6, 22828 (2016), Application(s): Fluorescence microscopy on human colorectal adenocarcinoma DLD1 cells, Abstract; Full Text
Low-level light in combination with metabolic modulators for effective therapy of injured brain: T. Dong, et al.; J. Cereb. Blood Flow Metab. 35, 1435 (2015), Application(s): Immunofluorescence Assay, Abstract; Full Text
Quantitative measurement of redox potential in hypoxic cells using SERS nanosensors: J. Jiang, et al.; Nanoscale 6, 12104 (2014), 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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