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ROS Brite™ HPF *Optimized for Detecting Reactive Oxygen Species (ROS)*

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Physical properties
Molecular weight424.40
Spectral properties
Excitation (nm)498
Emission (nm)517
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 have 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. ROS are also generated by exogenous sources such as ionizing radiation. Under conditions of oxidative stress, ROS production is dramatically increased, resulting in subsequent alteration of membrane lipids, proteins, and nucleic acids. Oxidative damage of these biomolecules is associated with aging as well as with a variety of pathological events, including atherosclerosis, carcinogenesis, ischemic reperfusion injury, and neurodegenerative disorders. ROS Brite™ HPF is a fluorogenic probe to measure hydroxyly radical in cells using conventional fluorescence microscopy, high-content imaging, microplate fluorometry, or flow cytometry. The cell-permeant ROS Brite™ HPF reagent is nonfluorescent and produces bright green fluorescence upon reaction with hydroxyl radical. The resulting fluorescence can be measured using fluorescence imaging, high-content imaging, microplate fluorometry, or flow cytometry. In the presence of peroxidase, HPF also reacts with hydrogen peroxide. HPF has good selectivity to hydroxyl radical compared to other ROS (e.g., supoeroxide and nitric oxide). APF and HPF show relatively high resistance to light-induced oxidation. APF and HPF are nonfluorescent until they react with the hydroxyl radical or peroxynitrite anion.


Flow cytometer

Excitation488 nm laser
Emission530/30 nm filter
Instrument specification(s)FITC channel

Fluorescence microscope

ExcitationFITC filter set
EmissionFITC filter set
Recommended plateBlack wall/clear bottom

Fluorescence microplate reader

Excitation490 nm
Emission525 nm
Cutoff515 nm
Recommended plateSolid black
Instrument specification(s)Bottom read mode


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of ROS Brite™ HPF *Optimized for Detecting Reactive Oxygen Species (ROS)* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM235.627 µL1.178 mL2.356 mL11.781 mL23.563 mL
5 mM47.125 µL235.627 µL471.254 µL2.356 mL4.713 mL
10 mM23.563 µL117.813 µL235.627 µL1.178 mL2.356 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

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Spectral properties

Excitation (nm)498
Emission (nm)517



View all 8 citations: Citation Explorer
Ultrasmall metal alloy nanozymes mimicking neutrophil enzymatic cascades for tumor catalytic therapy
Authors: Meng, Xiangqin and Fan, Huizhen and Chen, Lei and He, Jiuyang and Hong, Chaoyi and Xie, Jiaying and Hou, Yinyin and Wang, Kaidi and Gao, Xingfa and Gao, Lizeng and others,
Journal: Nature Communications (2024): 1626
Material-engineered bioartificial microorganisms enabling efficient scavenging of waterborne viruses
Authors: Li, Huixin and Xu, Yanpeng and Wang, Yang and Cui, Yihao and Lin, Jiake and Zhou, Yuemin and Tang, Shuling and Zhang, Ying and Hao, Haibin and Nie, Zihao and others,
Journal: Nature Communications (2023): 4658
Material-Engineered Bioartificial Microorganisms Enable Efficient Waterborne Viruses Scavenging
Authors: Li, Huixin and Cui, Yihao and Lin, Jiake and Zhang, Ying and Hao, Haibin and Wang, Yang and Xu, Yan and Nie, Zi-Hao and Zhou, Yuemin and Wang, Xiao-Yu and others,
Journal: (2022)
Toxin-Enabled “On-Demand” Liposomes for Enhanced Phototherapy to Treat and Protect against Methicillin-Resistant Staphylococcus aureus Infection
Authors: Zhuge, Deli and Chen, Mengchun and Yang, Xuewei and Zhang, Xufei and Yao, Lulu and Li, Li and Wang, Haonan and Chen, Hao and Yin, Qingqing and Tian, Dongyan and others,
Journal: Small (2022): 2203292
Molecular Hydrogen Attenuates High Hydrostatic Pressure-Induced Neuronal Cell Damage by Reversing Dysfunction of Mitochondrial Electron Transfer Chain
Authors: Lu, Zhuoyang and Zhang, Tiantian and Hu, Yachong and Liu, Hui and Cui, Li and Long, Jiangang and Liu, Jiankang
Journal: (2021)
Full-Process Radiosensitization Based on Nanoscale Metal--Organic Frameworks
Authors: Gong, Teng and Li, Yanli and Lv, Bin and Wang, Han and Liu, Yanyan and Yang, Wei and Wu, Yelin and Jiang, Xingwu and Gao, Hongbo and Zheng, Xiangpeng and others,
Journal: ACS nano (2020): 3032--3040
Bifunctionalized Novel Co-V MMO Nanowires: Intrinsic Oxidase and Peroxidase Like Catalytic Activities for Antibacterial Application
Authors: Wang, Yi and Chen, Chao and Zhang, Dun and Wang, Jin
Journal: Applied Catalysis B: Environmental (2019): 118256
Oxidative Stress--An Update and Insight in the Romanian Family Physician’s Adoption of the Concept
Authors: Berghea, Florian and Berghea, Camelia Elena and Abobului, Mihai
Journal: Internal Medicine : 11--15


View all 22 references: Citation Explorer
Developmental toxicity evaluation of three hexabromocyclododecane diastereoisomers on zebrafish embryos
Authors: Du M, Zhang D, Yan C, Zhang X.
Journal: Aquat Toxicol (2012): 1
MAPK inhibitors and siRNAs differentially affect cell death and ROS levels in arsenic trioxide-treated human pulmonary fibroblast cells
Authors: Park WH., undefined
Journal: Oncol Rep (2012): 1611
MG132, a proteasome inhibitor, induces human pulmonary fibroblast cell death via increasing ROS levels and GSH depletion
Authors: Park WH, Kim SH.
Journal: Oncol Rep (2012): 1284
Gallic acid-induced lung cancer cell death is accompanied by ROS increase and glutathione depletion
Authors: You BR, Kim SZ, Kim SH, Park WH.
Journal: Mol Cell Biochem (2011): 295
Proteasome inhibition by MG132 induces growth inhibition and death of human pulmonary fibroblast cells in a caspase-independent manner
Authors: You BR, Park WH.
Journal: Oncol Rep (2011): 1705
MAPK inhibitors differentially affect gallic acid-induced human pulmonary fibroblast cell growth inhibition
Authors: Park WH., undefined
Journal: Mol Med Report (2011): 193
Enhancement of gallic acid-induced human pulmonary fibroblast cell death by N-acetyl cysteine and L-buthionine sulfoximine
Authors: You BR, Park WH.
Journal: Hum Exp Toxicol (2011): 992
beta-Lapachone induces heart morphogenetic and functional defects by promoting the death of erythrocytes and the endocardium in zebrafish embryos
Authors: Wu YT, Lin CY, Tsai MY, Chen YH, Lu YF, Huang CJ, Cheng CM, Hwang SP.
Journal: J Biomed Sci (2011): 70
Mitogen-activated protein kinase inhibitors differently affect the growth inhibition and death of a proteasome inhibitor, MG132-treated human pulmonary fibroblast cells
Authors: Park WH., undefined
Journal: Hum Exp Toxicol (2011): 1945
Hydrogen protects vestibular hair cells from free radicals
Authors: Taura A, Kikkawa YS, Nakagawa T, Ito J.
Journal: Acta Otolaryngol Suppl (2010): 95