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

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
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Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	N/A
Solvent	DMSO

Spectral properties

Excitation (nm)	500
Emission (nm)	582

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	12352200

Overview SDS Protocol

Example protocol

PREPARATION OF STOCK SOLUTIONS

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)

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.

PREPARATION OF WORKING SOLUTION

MitoROS™ 580 Working Solution(2X)

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.

SAMPLE EXPERIMENTAL PROTOCOL

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.

Treat cells as desired.
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.
Wash cells gently three times and replace it with HHBS buffer.
Analyze the cells with a proper fluorescence instrument (e.g., a fluorescence microscope, flow cytometer) with Ex/Em = 510/580 nm.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)	500
Emission (nm)	582

Images

Figure 1. 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.

Citations

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)

Actives from the Micro-Immunotherapy Medicine 2LMIREG{\textregistered} Reduce the Expression of Cytokines and Immune-Related Markers Including Interleukin-2 and HLA-II While Modulating Oxidative Stress and Mitochondrial Function
Authors: Jacques, Camille and Marchand, Flora and Chatelais, Mathias and Floris, Ilaria
Journal: Journal of Inflammation Research (2024): 1161--1181

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

References

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
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Association rule mining of cellular responses induced by metal and metal oxide nanoparticles
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Journal: Analyst (2014): 943

Muscadine grape skin extract reverts snail-mediated epithelial mesenchymal transition via superoxide species in human prostate cancer cells
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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