Hi Numan

We used H2O2 with success

Effects of H2O2 exposure on human sperm motility parameters, reactive oxygen species levels and nitric oxide levels

SS Du Plessis, DA McAllister, A Luu, J Savia, A Agarwal, F Lampiao

Andrologia 42 (3), 206-210

Thanks Stefan S Du Plessis for the suggestion. I also wish to use H2O2 at a concentration of 200 micro-mole.

H2O2 would be a good and cheap reagent but it is unstable and easily neutralizes by cells. Instead cumene hydroperoxide would be a worthy replacement.

The concentration is dependent of cell type and the method. 1 mM would induce ROS in a short time (1 hour).

@Farjam Goudarzi can you send me any paper related to this approach. By the way I just want to induce stress and for that H2O2 is ok. Don't you think so

https://www.jstage.jst.go.jp/article/expanim/62/1/62_12-0037/_article

H2O2 is good but unstable. You can use Luperox® TBH70X, tert-Butyl hydroperoxide solution 70% in H2O (Sigma-Aldrich, catalog number: 458139). It is also cheap, and has an advantage over hydrogen peroxide in that it is less labile.

1) Article Oxidative Stress Assays (arsenite and tBHP) in Caenorhabditis elegans

2) https://www.nature.com/articles/s41598-018-27614-8

Thanks @ Frederic Nduhirabandi I will look into these articles you recommended

If you are just intended to induce ROS, H2O2 will be sufficed for that purpose, dont bother.

Hi Numan

You should check further with Farjam. He recently published a paper with the same method using H2O2 in stem cells you intend to use. But to my experience the only problem is that it is less stable, and you may experience inconsistencies in your data depending on the concentration you use and the level of injury/cell death. Please check additional answers on the issues of H2O2 at https://www.researchgate.net/search.Search.html?type=question&query=H2O2.

All the best!

Thanks Frederic and Farjam for your valuable comments.