ROS Brite™ 670 *Optimized for Detecting Reactive Oxygen Species (ROS)*

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1 mg 16002 $195

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Telephone: 1-800-990-8053
Fax: 1-408-733-1304
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Ex/Em (nm)650/669
Storage Freeze (<-15 °C)
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Category Neurobiology
Reactive Oxygen Species
Related Cell Signaling
Secondary Reagents
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™ 670 reagent is a new fluorogenic probe to measure oxidative stress in cells using conventional fluorescence microscopy, high-content imaging, microplate fluorometry, or flow cytometry. The cell-permeant ROS Brite™ 670 reagent is nonfluorescent and produces bright near-infrared fluorescence upon ROS oxidation. The resulting fluorescence can be measured using fluorescence imaging, high-content imaging, microplate fluorometry, or flow cytometry.

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of ROS Brite™ 670 *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.

Molarity calculator

Table 2. Enter any two values (mass, volume, concentration) to calculate the third.

Mass Molecular weight Volume Concentration Moles
/ = x =

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This protocol only provides a guideline, and should be modified according to your specific needs.

1)       Prepare a 10 to 20 mM ROS Brite™ stock solution in DMSO. Make 5 to 10 µM working solution by diluting the DMSO stock solution into Hanks solution with 20 mM Hepes buffer (HHBS).

2)       Treat cells as desired (e.g., RASM cells are treated with 50-100 nM angiotensin II for 3-5 hours).

3)       Incubate the cells with ROS Brite™ (5-10 µM, from Step #1) for 15 -30 minutes at 37 ºC.

Note: The concentration of ROS Brite™ used varies with different cell lines, one will need test with different concentrations to get the optimal dose. For cat#16053, one might use less concentration such as 0.5-5 uM.

4)       Replace the dye-loading solution with HHBS buffer.

5)       Analyze the cells with a proper fluorescence instrument (e.g., a fluorescence microscope, flow cytometer or an in vivo imaging instrument).


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Safety Data Sheet (SDS)

1. Reactive Oxygen Species (ROS) Detection

Application Notes
1. AssayWise Letters 2013, Vol 2(1)

Certificate of Analysis