Intracellular Total ROS Activity Assay

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The Intracellular Total ROS Activity Assay enables the screening of oxidative stress inhibitor and activator reagents for potency. The assay assesses the overall level of intracellular ROS activity. Analyze using a flow cytometer.

Figure 1. Jurkat cells were stained with the Intracellular Total ROS Green dye (cat. 9144), then treated with a negative control (left) or 100 µM tert-Butyl hydroperoxide (ROS-inducing agent) (middle). The median fluorescence intensity (MFI) of stained negative control cells was 11,875 in FL1-A (Negative) and 419,067 for ROS-induced cells (Positive). The data are also shown in overlay (black: Negative, red: Positive). Data courtesy of Dr. Kristi Strandberg, ICT, 224:76.
Figure 1. Jurkat cells were stained with the Intracellular Total ROS Green dye (cat. 9144), then treated with a negative control (left) or 100 µM tert-Butyl hydroperoxide (ROS-inducing agent) (middle). The median fluorescence intensity (MFI) of stained negative control cells was 11,875 in FL1-A (Negative) and 419,067 for ROS-induced cells (Positive). The data are also shown in overlay (black: Negative, red: Positive). Data courtesy of Dr. Kristi Strandberg, ICT, 224:76.
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Figure 1. Jurkat cells were stained with the Intracellular Total ROS Green dye (cat. 9144), then treated with a negative control (left) or 100 µM tert-Butyl hydroperoxide (ROS-inducing agent) (middle). The median fluorescence intensity (MFI) of stained negative control cells was 11,875 in FL1-A (Negative) and 419,067 for ROS-induced cells (Positive). The data are also shown in overlay (black: Negative, red: Positive). Data courtesy of Dr. Kristi Strandberg, ICT, 224:76.

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Product Manual
Safety Data Sheet

SKU: 9144

Size: 100 Tests
Price:
$352.00
Ships: 1-2 business days
  1. Prepare samples and controls in 1X Assay Buffer or buffer of choice (0.5 mL at a cell density of 5 x 105 – 1 x 106 cells/mL).
  2. Reconstitute Total ROS Green with 100 µL DMSO to produce a 500X stock concentrate.
  3. Dilute 1:10 by adding 900 µL 1X Assay Buffer to the 500X stock Total ROS Green to produce a 50X working solution.
  4. Add Total ROS Green to each sample at 1:50. For example, add 10 µL into 490 µL cultured cells.
  5. Incubate 1 hour at 37°C.
  6. Treat cells with test compounds for desired period of time. Keep cells protected from light.
  7. Analyze with a flow cytometer. Total ROS Green excites at 490 nm and emits at 520 nm.
  • Total ROS Green, lyophilized, 1 vial, #6688
  • DMSO, 200 µL, 1 vial, #6689
  • Assay Buffer, 10X, 15 mL, #686
  • Kit Manual

    • Product Specific References

    PMID Publication
    38871787Wnuk, E, et al. 2024. The physiological levels of epigallocatechin gallate (EGCG) enhance the Cd-induced oxidative stress and apoptosis in CHO-K1 cells. Scientific reports, 13625.
    38806527Gassib, N., et al. 2024. Cellular mechanisms mediating the anti-cancer effects of carnosol on gingiva carcinoma. Scientific reports, 12266.

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