Cell Meter™ Fluorimetric Cellular Lipid Peroxidation Assay Kit

Protocol summary

1. Plate cells at desired confluency.
2. Treat cells with compound of interest and incubate.
3. Add Lipoxite™ R590/G525.
4. Remove media, and wash with PBS.
5. Analyze sample by fluorescence microscope or flow cytometer through FITC/TRITC or FITC/PE channels.

Important

Thaw one of each kit component at room temperature before starting the experiment.

Prepare 10X working solution of Lipoxite™ R590/G525 by making 1:50 dilution of 500X Lipoxite™ R590/G525 (Component A) into HHBS (Component B)

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

1. Grow cells at desired density and incubate overnight in a humidified chamber at 37°C with 5% CO₂.
   
   
2. Treat cells with test compounds as desired. Note: For a positive control, add hydrogen peroxide (Component C) to the cells at a final concentration of ~250 µM (1X) for 30 minutes.
   
   
3. Add 10X Lipoxite™ R590/G525 to the cells at a final concentration of 1X (for example add 10 uL to 90 uL of the cells).
   
   
4. Incubate the cells for 30 min at 37°C with 5% CO₂ cell incubator.
   
   
5. Remove media and wash cells with HHBS (Component B) or DPBS for three times.
   
   
6. Monitor fluorescence of cells with a fluorescence microscope or flow cytometer through FITC/TRITC or FITC/PE channels within 2 hours of staining.