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.
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
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).
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.
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.
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.
Detailed instructions are included in the manual for microscopy and flow cytometry applications for adherent and suspension cells.
Regulatory Status:
RUO - Research Use Only
Product Literature References
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Combinational phototherapy and hypoxia-activated chemotherapy favoring antitumor immune responses: B. Ma, et al.; Int. J. Nanomedicine 14, 4541 (2019), Application(s): Fluorescence microscopy using 4T1 cells, Abstract; Full Text
Di-(2-ethylhexyl) phthalate (DEHP) inhibits steroidogenesis and induces mitochondria-ROS mediated apoptosis in rat ovarian granulosa cells: A. Tripathi, et al.; Toxicol. Res. (Camb.) 8, 381 (2019), Application(s): Flow cytometry using granulosa cells, Abstract;
Encircling granulosa cells protects against di-(2-ethylhexyl)phthalate-induced apoptosis in rat oocytes cultured in vitro: A. Tripathi, et al.; Zygote 27, 203 (2019), Abstract;
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Oral ferroportin inhibitor ameliorates ineffective erythropoiesis in a model of β-thalassemia: V. Manolova, et al.; J. Clin. Invest. 130, 491 (2019), Application(s): Flow cytometry analysis using mouse red blood cells, Abstract;
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Solid matrix-supported supercritical CO2 enhances extraction of γ-linolenic acid from the cyanobacterium Arthrospira (Spirulina) platensis and bioactivity evaluation: X. Yang, et al.; Mar. Drugs 17, 203 (2019), Application(s): Fluorescence microscopy using Zebrafish larvae, Abstract; Full Text
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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
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General Literature References
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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;
Reagent used in high-throughput assays to monitor mitochondrial function and respiration rates, through the kinetic measurement of extracellular oxygen consumption.
A pH sensitive phosphorescent probe that can monitor cellular lactic acid extrusion, resulting from glycolysis, as a measure of glycolytic flux (ECA/ECAR)
