I am trying to detect ROS in some breast cancer cell lines, including MDA-MB-231, MCF-7 and 4T1. I found out that ROS in some cell lines decreased after the incubation with H2O2, while in some cell lines ROS increased. I used DCFH-DA as prober. I just feel confused about the results. Could someone provide some clues?
Dear Lan,
DCF shows a slight tendency to detect H2O2 better than other ROS supspecies, but it is not a probe that you can use to differ between different ROS subspecies.
Moreover, H2O2 itself is a ROS supecies. If you put H2O2 on your cells then you treat you cells with a substance you want to detect. H2O2 is a ROS subspecies.
It is basically wrong to say "I treated cells with H2O2 and looked at ROS production", since H2O2 istelf counts as ROS. It is the same as one would say "I put strawberrys in my jelly and analyzed, if strawberrys are in my jelly" ;) .
There are different ROS subspecies, such as superoxide, H2O2, and peroxyl- or hydroxyl radicals, but all of them are termed ROS. For example, IL-6, TNF and IL-1ß are all cytokines, but each works and is produced in very different ways. And of course, you need different detection ELISAs to analyze them.
If you like, please have a look at our recent review. It explains in detail what ROS are and which probes you can use do detect different ROS subspecies.
Article Functions of ROS in Macrophages and Antimicrobial Immunity
Please feel free to ask any time, if you have further questions.
All the best and stay healthy,
Marc
I have worked on the effect of some extracts on the inhibition of ROS, similar to what you mention, in my case, the extract could inhibit ROS compared to the sample that was not treated with the extract, i.e. it could decrease the oxidative stress in the cell. But I have also read articles in which they work with cell lines, in which they try to provoke this oxidative stress in the cells to induce apoptosis. But as mentioned in the previous answer, it is not possible to know what specific ROS you are measuring.
I think it will be helpful for you go through this link
https://www.thermofisher.com/np/en/home/life-science/cell-analysis/cell-viability-and-regulation/oxidative-stress.html?ef_id=Cj0KCQjw5PGFBhC2ARIsAIFIMNdSiE5gT_SpxP-xNo-5svwszyncML4_DZAchhgk9hA0Hx2vY94_mOkaAnzzEALw_wcB:G:s&s_kwcid=AL!3652!3!378136343458!b!!g!!%2Bros%20%2Bdetection&cid=bid_pca_iva_r01_co_cp1359_pjt0000_bid00000_0se_gaw_nt_pur_con&gclid=Cj0KCQjw5PGFBhC2ARIsAIFIMNdSiE5gT_SpxP-xNo-5svwszyncML4_DZAchhgk9hA0Hx2vY94_mOkaAnzzEALw_wcB
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