Intracellular Reactive Oxygen Species (ROS) Detection Assay Kit

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Size	Unit Price	Philadelphia, PA	Portland, OR	Japan*	Quantity
1KIT	$217.00	Contact Us	Contact Us	28

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Product Information

• Size : 96 tests

Description

• Reactive Oxygen Species (ROS) are mainly produced by mitochondria, regulating processes necessary for cell proliferation, including in cancer cells. Cell death, such as apoptosis and ferroptosis, is associated with an increased intracellular ROS level.¹⁾ DCFH-DA is enzymatically deacetylated and hydrolyzed to produce DCFH, which penetrates the cell. Intracellular DCFH is then oxidized to DCF by reactive oxygen species (e.g., OH, O₂^-, H₂O₂, etc.), resulting in a strong fluorophoric response that turns the solution greenish-yellow in color.²⁾ This kit is designed to directly visualize the real-time production of activated oxygen species in live cells using fluorescence microscopy equipped with a standard green filter set (450-490 nm / 500-550 nm), or flow cytometry equipped with a blue laser (488 nm / 520 nm).

Components

• 1000X Reaction Dye (DCFH-DA) : 10 μL
• 10X Reaction Buffer : 25 mL×2

Preparation

• 1X Reaction Buffer : Reconstitute the 10X Reaction Buffer by distilled water.
• 1X Reaction Dye : Reconstitute the 1000X Reaction Dye by 1X Reaction Buffer.

Step–by–Step Protocol

• See the pdf file in this link.

Storage

• Stock the 10X Reaction Buffer and 1X Reaction Buffer at 4 °C. Stock the 1000X Reaction dye at -20 °C. Immediately use up the 1X Reaction Dye after reconstitution.

References

1. 1) Mitochondrial ROS regulation of proliferating cells
   • L. Diebold, N. S. Chandel, Free Radic. Biol. Med. 2016, 100, 86.
2. 2) Time-dependent interactions of oxidant-sensitive fluoroprobes with inhibitors of cellular metabolism
   • K. E Tollefson, J. Kroczynski, M. V Cutaia, Lab. Invest. 2003, 83, 367.