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Neurl (BC058386) Mouse Untagged Clone

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Cat# MC218205

Size : 10µg

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

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

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

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)

  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.

PREPARATION OF WORKING SOLUTION

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.

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.

  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.

Spectrum

Citations

View all 37 citations: Citation Explorer
Mitochondria-targeted nanovesicles for ursodeoxycholic acid delivery to combat neurodegeneration by ameliorating mitochondrial dysfunction
Authors: Zhang, Shizheng and Li, Mengmeng and Li, Yuan and Yang, Shike and Wang, Jian and Ren, Xiaoxiang and Wang, Xiuhui and Bai, Long and Huang, Jianping and Geng, Zhen and others,
Journal: Journal of Nanobiotechnology (2025): 202
Anti-Cancer and Pro-Immune Effects of Lauric Acid on Colorectal Cancer Cells
Authors: Mori, Shiori and Fujiwara-Tani, Rina and Ogata, Ruiko and Ohmori, Hitoshi and Fujii, Kiyomu and Luo, Yi and Sasaki, Takamitsu and Nishiguchi, Yukiko and Bhawal, Ujjal Kumar and Kishi, Shingo and others,
Journal: International Journal of Molecular Sciences (2025): 1953
Sevoflurane exposure accelerates the onset of cognitive impairment via promoting p-Drp1S616-mediated mitochondrial fission in a mouse model of Alzheimer's disease
Authors: He, Kaiwu and Li, Youzhi and Xiong, Wei and Xing, Yanmei and Gao, Wenli and Du, Yuting and Kong, Wei and Chen, Lixin and Yang, Xifei and Dai, Zhongliang
Journal: Free Radical Biology and Medicine (2024)
Photocatalytic scaffolds enhance anticancer performances of bacterial consortium AUN
Authors: Miyahara, Mikako and Doi, Yuki and Takaya, Naoki and Miyako, Eijiro
Journal: Chemical Engineering Journal (2024): 156378
Gastrodin attenuates high fructose-induced sweet taste preference decrease by inhibiting hippocampal neural stem cell ferroptosis
Authors: Tang, Chuan-Feng and Ding, Hong and Wu, Ya-Qian and Miao, Zi-An and Wang, Zi-Xuan and Wang, Wen-Xuan and Pan, Ying and Kong, Ling-Dong
Journal: Journal of Advanced Research (2024)

References

View all 46 references: Citation Explorer
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
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
Page updated on April 15, 2025
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