I have searched some research articles related to Ferroptosis. Many kinds of experiments have been done. like, staining, GSH assay, lipid peroxidation assay, etc.
So,
1. What methods (both basic and confirmatory) should be followed?
2. Most of the researchers have been used in the kit. Any method is available other than using the kit?
Ferroptosis is driven by unchecked/lethal accumulation of lipid peroxides in plasma membranes as a result of failed glutathione-dependent antioxidant defenses, leading to cell death. That said, use of ferroptosis-specific antibody would facilitate examining. One can use ferroptosis specific inhibitors (ferrostatin-, liproxstatin-1 and vitamin E) or classic ferroptosis inducer (Erastin) as positive controls.
I did come across the below detailed method and may be of any use for you
https://link.springer.com/protocol/10.1007%2F978-1-0716-0247-8_11
Hi Essha Chatterjee
In general, evaluation of lipid ROS in cells is the most common way for ferroptosis detection. For this purpose, you can use Liperfluo, C11-BODIPY 581/591, or MDA assay kits. Moreover, you can assay cell viability after using ferroptosis inhibitors, such as ferrostatin-1, liproxstatin, and α-Tocopherol to confirm the occurrence of ferroptosis.
Based on your research you can also assay GSH or iron levels, which is usually optional.
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