text.skipToContent text.skipToNavigation

Maximum quantity allowed is 999

Please select the quantity

Intracellular Reactive Oxygen Species (ROS) Detection Assay Kit

Intracellular reactive oxygen species (ROS) are mainly produced in mitochondria. ROS react to lipids or proteins inside cells may inhibit the cell functions and induce a series of pathological processes including cell death, aging and oncogenesis.
The Intracellular Reactive Oxygen Species (ROS) Detection Assay Kit provides a fluorescence probe, DCFH-DA, suitable for immediate detection of ROS. This kit is sufficient for one 96-well microplate assay and requires 1 mL of test solution for 1 × 106 cells/mL. The optimal working concentration of the reaction dye may vary depending on the cell lines being used.


Figure. Intracellular generation of reactive oxygen species

Kit Components

  • 1000×Reaction Dye (2',7'-Dichlorofluorescein Diacetate) : 10 μL for 100 tests
  • 10×Reaction Buffer : 25 mL×2 for 100 tests

Page Top

Application 1: Fluorescence observation of ROS produced in Hela cells

  1. Culture the HeLa cells in 96 well culture plate and grow to 80% confluent.
  2. Remove the culture media. Treat the cells with 1 × Reaction Dye and incubate for 30 minutes in CO2 incubator (37°C).
  3. Observe the Intracellular ROS (green fluorescence) by fluorescence microscope.

Fluorescence microscopy observation of ROS produced in Hela cells

Figure 1. Fluorescence microscopy observation of ROS produced in Hela cells
  ROS produced in HeLa cells were observed by fluorescence microscope.

Page Top

Application 2-1: Comparison of ROS produced in NIH-3T3 treated with a ROS inducer and a ROS inhibitor

  1. Culture the NIH-3T3 cells in 96 well culture plate and grow to 80% confluent.
  2. Remove the culture media from a sample of cells and treat cells with a ROS inducer and incubate in a CO2 incubator (37°C) overnight.
  3. Remove the ROS inducer from a sample of cells and additionally treat cells with a ROS inhibitor and incubate in a CO2 incubator (37°C) for 30 minutes.
  4. Treat all cells with 1 × Reaction Dye in a CO2 incubator (37°C) for 30 minutes.
  5. Test intracellular ROS production on a plate reader.

Detection of ROS in NIH-3T3 cells using a plate reader

Conditions for addition of ROS accelerators and ROS inhibitors

Figure 2. Detection of ROS in NIH-3T3 cells using a plate reader
  ROS production was induced by a ROS inducer and inhibited by a ROS inhibitor.

Page Top

Application 2-2: ROS analyzed in NIH-3T3 with and without a ROS inducer by flow cytometry

  1. Culture the NIH-3T3 at 105 cells/mL and grow to 80% confluent.
  2. Remove the culture media from a sample of cells and treat cells with 1 mM Erastin and incubate in a CO2 incubator (37°C) overnight.
  3. Collect the cells and treat cells with 1 × Reaction Dye in a CO2 incubator (37°C) for 30 minutes.
  4. Analyze the intracellular ROS production by flow cytometry.

Without Treatment
Detection of ROS in NIH-3T3 cells not treated with a ROS inducer by flow cytometer
With Treatment
Detection of ROS in NIH-3T3 cells treated with a ROS inducer by flow cytometer

Figure 3. Detection of ROS in NIH-3T3 cells by flow cytometer
  ROS production was induced by Erastin treatment.

Page Top

Session Status
Your session will timeout in 10 minutes. You will be redirected to the HOME page after session timeout. Please click the button to continue session from the same page. minute. You will be redirected to the HOME page after session timeout. Please click the button to continue session from the same page.

Your session has timed out. You will be redirected to the HOME page.