Intracellular Total ROS Activity Assay

SKU: orb1409908

Description

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.

Research Area

Signal Transduction

Images & Validation

• Application Notes:
  1. Prepare samples and controls. 2. Dilute 10X Cellular Assay Buffer 1:10 with diH2O. 3. Reconstitute FAM-FLICA with 50 µL DMSO. 4. Dilute FAM-FLICA 1:5 by adding 200 µL PBS. 5. Add diluted FAM-FLICA to each sample at 1:30-1:60 (e.g. spike at 1:30 by adding 10 µL to 290 µL sample). 6. Incubate approximately 1 hour. 7. Remove media and wash cells 3 times: add 1X Cellular Wash Buffer and spin cells. 8. If desired, label with additional stains, such as Hoechst 33342, Propidium Iodide, 7-AAD, or an antibody. 9. If desired, fix cells. 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM-FLICA excited at 492 nm and emits at 520 nm.

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Key Properties

• Sample Types: Cell culture

Storage & Handling

• Storage: Total ROS Green and DMSO at ≤-20°C, Assay Buffer at ≤2-8°C
• Expiration Date: 6 months from date of receipt.
• Disclaimer: For research use only