Intracellular Total ROS Activity Assay

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.
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:
$359.75
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

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