AAT Bioquest

MitoROS™ 580 *Optimized for Detecting Reactive Oxygen Species (ROS) in Mitochondria*

Fluorescence response of MitoROS™ 580 (10 µM)to different reactive oxygen species (ROS) and reactive nitrogen species (RNS). The fluorescence intensities were monitored at Ex/Em = 540/590 nm.
Fluorescence response of MitoROS™ 580 (10 µM)to different reactive oxygen species (ROS) and reactive nitrogen species (RNS). The fluorescence intensities were monitored at Ex/Em = 540/590 nm.
Fluorescence response of MitoROS™ 580 (10 µM)to different reactive oxygen species (ROS) and reactive nitrogen species (RNS). The fluorescence intensities were monitored at Ex/Em = 540/590 nm.
Ordering information
Catalog Number
Unit Size
Add to cart
Additional ordering information
InternationalSee distributors
Bulk requestInquire
Custom sizeInquire
ShippingStandard overnight for United States, inquire for international
Request quotation
Physical properties
Molecular weightN/A
Spectral properties
Excitation (nm)500
Emission (nm)582
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure


Molecular weight
Excitation (nm)
Emission (nm)
Reactive oxygen species (ROS) are chemically reactive molecules containing oxygen. Examples include superoxide, hydroxyl radical, singlet oxygen, and peroxides. ROS is highly reactive due to the presence of unpaired valence shell electrons. ROS forms as a natural byproduct of the normal metabolism of oxygen and has important roles in cell signaling and homeostasis. However, during times of environmental stress (e.g., UV or heat exposure), ROS levels can increase dramatically. This may result in significant damage to cell structures. Cumulatively, this is known as oxidative stress. MitoROS™ 580 is a superoxide-sensitive dye that is localized in mitochondria upon loading into live cells. Oxidation of MitoROS™ 580 by superoxide generates red fluorescence. MitoROS™ 580 can be used to monitor superoxide in mitochondria either with a fluorescence microscope or flow cytometer. MitoROS™ 580 reagent permeates live cells where it selectively targets mitochondria. It is rapidly oxidized by superoxide. It is less likely to be oxidized by other reactive oxygen species (ROS) and reactive nitrogen species (RNS). The oxidized product is highly fluorescent in cells. MitoROS™ 580 provides a valuable tool for investigating oxidative stress in various pathologies. MitoROS™ 580 is equivalent to the MitoSOX™ Mitochondrial Superoxide Indicator (#M36008) that is often used for live-cell imaging (MitoSOX™ is the trademark of ThermoFisher).

Example protocol


Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles

MitoROS™ 580 Stock Solution (1000X)
  1. Add 13 µL of DMSO to the MitoROS™ 580 vial and mix well.

    Note: Any unused stock solution can be stored at -20 °C, protected from light.


MitoROS™ 580 Working Solution(2X)
  1. Dilute the DMSO stock solution into Hanks solution with 20 mM Hepes buffer (HHBS) to make a 2X working solution.

    Note: The 2X MitoROS™ 580 working solution is not stable, use it promptly.


Important Note

This protocol is a mere guideline and should be optimized to suit your specific requirements. Prior to making the MitoROS™ 580 working solution, treat cells as desired.

  1. Treat cells as desired.

  2. Incubate the cells (such as 100 µL/well in 96-well plate) with equal volume of 2X MitoROS™ 580 working solution for 10-30 minutes at 37 °C, protected from light.

    Note: The final in-cell concentration of the MitoROS™ 580 should not exceed 1X. Higher concentrations can lead to cytotoxic effects, such as altered mitochondrial morphology and fluorescence redistribution to nuclei and cytosol.

    Note: Different cells react to MitoROS™ 580 differently, adjust the working concentration accordingly.

  3. Wash cells gently three times and replace it with HHBS buffer.

  4. Analyze the cells with a proper fluorescence instrument (e.g., a fluorescence microscope, flow cytometer) with Ex/Em = 510/580 nm.


Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)500
Emission (nm)582



View all 27 citations: Citation Explorer
ARK5 enhances cell survival associated with mitochondrial morphological dynamics from fusion to fission in human multiple myeloma cells
Authors: Karnan, Sivasundaram and Hanamura, Ichiro and Ota, Akinobu and Vu, Lam Quang and Uchino, Kaori and Horio, Tomohiro and Murakami, Satsuki and Mizuno, Shohei and Rahman, Md Lutfur and Wahiduzzaman, Md and others,
Journal: Cell Death Discovery (2024): 56
Claudin-1 enhances chemoresistance of human lung adenocarcinoma A549 cells mediated by forming an amino acid barrier
Authors: Kimura, Riho and Ito, Ayaka and Hashimoto, Shotaro and Eguchi, Hiroaki and Nasako, Haruka and Takashina, Yui and Morikawa, Yoshifumi and Suenami, Koichi and Yoshino, Yuta and Endo, Satoshi and others,
Journal: (2024)
Oxidized-LDL Induces Metabolic Dysfunction in Retinal Pigment Epithelial Cells
Authors: Tomomatsu, Manami and Imamura, Naoto and Izumi, Hoshimi and Watanabe, Masatsugu and Ikeda, Masataka and Ide, Tomomi and Uchinomiya, Shohei and Ojida, Akio and Jutanom, Mirinthorn and Morimoto, Kazushi and others,
Journal: Biological and Pharmaceutical Bulletin (2024): 641--651
Melanin-like nanoparticles alleviate ischemia-reperfusion injury in the kidney by scavenging reactive oxygen species and suppressing ferroptosis
Authors: Feng, Wenxiang and Zhu, Nan and Xia, Yubin and Huang, Zehai and Hu, Jianmin and Guo, Zefeng and Li, Yuzhu and Zhou, Song and Liu, Yongguang and Liu, Ding
Journal: iScience (2024)
Quercetin Impairs the Growth of Uveal Melanoma Cells by Interfering with Glucose Uptake and Metabolism
Authors: Tura, Ayseg{\"u}l and Herfs, Viktoria and Maa{\ss}en, Tjorge and Zuo, Huaxin and Vardanyan, Siranush and Prasuhn, Michelle and Ranjbar, Mahdy and Kakkassery, Vinodh and Grisanti, Salvatore
Journal: International Journal of Molecular Sciences (2024): 4292
Curcumin/L-OHP co-loaded HAP for cGAS-STING pathway activation to enhance the natural immune response in colorectal cancer
Authors: Xiao, Yao and Guo, Guohu and Wang, Huaiming and Peng, Bin and Lin, Yinglin and Qu, Gaowen and Li, Ben and Jiang, Zhaojun and Zhang, Fan and Wu, Jiaming and others,
Journal: Bioengineering \& Translational Medicine (2023): e10610
Identification of Lipocalin 2 as a Ferroptosis-Related Key Gene Associated with Hypoxic-Ischemic Brain Damage via STAT3/NF-$\kappa$B Signaling Pathway
Authors: Luo, Lianxiang and Deng, Liyan and Chen, Yongtong and Ding, Rui and Li, Xiaoling
Journal: Antioxidants (2023): 186
Berberine Induces Combined Cell Death in Gastrointestinal Cell Lines
Authors: Mori, Shiori and Fujiwara-Tani, Rina and Gyoten, Momoko and Nukaga, Shota and Sasaki, Rika and Ikemoto, Ayaka and Ogata, Ruiko and Kishi, Shingo and Fujii, Kiyomu and Kuniyasu, Hiroki
Journal: International Journal of Molecular Sciences (2023): 6588
Inhibiting mir-34a-5p regulates doxorubicin-induced autophagy disorder and alleviates myocardial pyroptosis by targeting Sirt3-AMPK pathway
Authors: Zhong, Zuoquan and Gao, Yefei and Zhou, Jiedong and Wang, Fang and Zhang, Peipei and Hu, Songqing and Wu, Haowei and Lou, Haifei and Chi, Jufang and Lin, Hui and others,
Journal: Biomedicine \& Pharmacotherapy (2023): 115654


View all 46 references: Citation Explorer
Nonthermal plasma induces head and neck cancer cell death: the potential involvement of mitogen-activated protein kinase-dependent mitochondrial reactive oxygen species
Authors: Kang SU, Cho JH, Chang JW, Shin YS, Kim KI, Park JK, Yang SS, Lee JS, Moon E, Lee K, Kim CH.
Journal: Cell Death Dis (2014): e1056
An oxidative stress mechanism of shikonin in human glioma cells
Authors: Yang JT, Li ZL, Wu JY, Lu FJ, Chen CH.
Journal: PLoS One (2014): e94180
Low Amounts of Mitochondrial Reactive Oxygen Species Define Human Sperm Quality
Authors: Marques M, Sousa AP, Paiva A, Almeida-Santos T, Ramalho-Santos J.
Journal: Reproduction. (2014)
Subneurotoxic copper(II)-induced NF-kappaB-dependent microglial activation is associated with mitochondrial ROS
Authors: Hu Z, Yu F, Gong P, Qiu Y, Zhou W, Cui Y, Li J, Chen H.
Journal: Toxicol Appl Pharmacol (2014): 95
HPLC-based monitoring of products formed from hydroethidine-based fluorogenic probes--the ultimate approach for intra- and extracellular superoxide detection
Authors: Kalyanaraman B, Dranka BP, Hardy M, Michalski R, Zielonka J.
Journal: Biochim Biophys Acta (2014): 739
The acute inhibitory effect of iodide excess on sodium/iodide symporter expression and activity involves the PI3K/Akt signaling pathway
Authors: Serrano-Nascimento C, da Silva Teixeira S, Nicola JP, Nachbar RT, Masini-Repiso AM, Nunes MT.
Journal: Endocrinology (2014): 1145
Association rule mining of cellular responses induced by metal and metal oxide nanoparticles
Authors: Liu R, France B, George S, Rallo R, Zhang H, Xia T, Nel AE, Bradley K, Cohen Y.
Journal: Analyst (2014): 943
Muscadine grape skin extract reverts snail-mediated epithelial mesenchymal transition via superoxide species in human prostate cancer cells
Authors: Burton LJ, Barnett P, Smith B, Arnold RS, Hudson T, Kundu K, Murthy N, Odero-Marah VA.
Journal: BMC Complement Altern Med (2014): 97
The omega-3 polyunsaturated fatty acid DHA induces simultaneous apoptosis and autophagy via mitochondrial ROS-mediated Akt-mTOR signaling in prostate cancer cells expressing mutant p53
Authors: Shin S, Jing K, Jeong S, Kim N, Song KS, Heo JY, Park JH, Seo KS, Han J, Park JI, Kweon GR, Park SK, Wu T, Hwang BD, Lim K.
Journal: Biomed Res Int (2013): 568671
Moderate hypoxia followed by reoxygenation results in blood-brain barrier breakdown via oxidative stress-dependent tight-junction protein disruption
Authors: Zehendner CM, Librizzi L, Hedrich J, Bauer NM, Angamo EA, de Curtis M, Luhmann HJ.
Journal: PLoS One (2013): e82823