I have done some luminol based ROS assays, mainly using this protocol:
https://bio-protocol.org/e1685
However, I only get a ROS response using FLG22. I have also tried to expose my leaf discs to high salt (150mM), sorbitol (300mM) cold (putting the 96 well plates in ice water), but none have given any ROS response.
I was wondering if anybody has any idea why this doesn't work? Or how I could make this work. I'd prefer the luminol based assays. But I will probably start using different methods if is not possible to test abiotic stresses this way.
Max,
ROS are accumulated under abiotic stress conditions, which cause oxidative damage and eventually resulting in cell death. But, I guess that high salt, sorbitol, and cold could be not sufficient to start cell death in small discs at 96-well plates.
Let me clarify that I am interrested in the ROS pulse you get from the first sensing of a stress. Not the ROS that is produced when the cell is actually damaged, which is on longer time scales
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