I am planning to study oxidative stress in bacteria and protein damage via reactive oxygen species (ROS) by antibacterial agents.
Thanks a lot,
most.
Hi Mostafa,
Initially, you would need to think about what type of ROS you would be interested, as some of the employed assays would focus or be specific ("at least in theory") to one type of ROS (e.g. singlet oxygen, superoxide anion or hydrogen peroxide). Some of them can be quite broad and non-specific assays. I guess that the method you will choose depend mostly on the equipment available, as many of the are fluorescent and chemiluminescent.
In addition, as working with bacteria, you would not need to worry too much with compartimentalization of ROS production as it would be with eukaryotic cells. This could be quite tricky.
it would be better to read some reviews about this area, I would suggest the following two:
Wooley et al., 2014. Recent advances in reactive oxygen species measurement in biological systems
http://www.sciencedirect.com/science/article/pii/S0968000413001552
Dikalov et al., 2014. Methods for Detection of Mitochondrial and Cellular Reactive Oxygen Species
http://online.liebertpub.com/doi/full/10.1089/ars.2012.4886
I hope I could hell,
Cheers!
Hello, please refer to our recent publications enclosed herewith. I am sure, you shall get a comprehensive idea about use of various markers for monitoring ROS levels. Please feel free to write you us for any clarification you might like to have.
Best.
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I read with interest this question and everyone's good answers. It appears that most of the cited methods were developed for mammalian cells. Are there any specific techniques for bacteria or yeast cultures? Is mesuring extracellular ROS (in the growth medium) easier than intracellular ones?
I used ( Acrydine Orange / Ethidium bromide assay ) it simple and reliable methods by mixing the same volume from bacterial suspension with Acrydine Orange / Ethidium bromide. incubate at room temperature for 5 min. and then prepare slide smear from mixture leave to dry then examined using fluorescent microscope. survival bacteria were appear orange while dead bacteria take red color.
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