We have a good delivery system which passes cell membrane, we want to generate Reactive Oxygen or free radicals inside cytozol, how to do that? It can be either a single substance or a combination of two which will react after release from the carrier.
The combination of xanthine and xanthine oxidase is a common way to do it, and will generate superoxide, which is then converted endogenously to H2O2 - adding H2O2 straight off does not really mimic the intracellular generation of ROS because there is no superoxide
The combination of xanthine and xanthine oxidase is a common way to do it, and will generate superoxide, which is then converted endogenously to H2O2 - adding H2O2 straight off does not really mimic the intracellular generation of ROS because there is no superoxide
Great, thank You. But I was rather meaning the group of cells inside a bigger organism, mouse. We have a selective (more or less) transporter and we want to generate ROS inside the targeted group of cells. Big organic peroxide we are already trying, xantine should be another solution, are there any others?
You can induce ROS with things like elevated glucose or cytokines, but this would also set off other signalling pathways, not just exclusively ROS. You're better off with a pure enzyme/substrate duo, such as X/XO, that will produce only superoxide in the first instance. Some people simply add NADPH, but lots of enzymes use this as an electron donor and you'll have potentially a lot of different products therein. Tissue specific overexpression of Nox2, which produces superoxide and nothing else, is another route some take.
Electron transport inhibitors like Antimycin-A, Rotenone, cyanide or heavy metal ions (in particular ferrous/ferric ions etc.) can rapidly generate ROS in cells. Paraquat/methyl viologen also can rapidly promote ROS generation.
Hi Tomasz!
Try pyocyanin and measure ROS as we did (http://www.nature.com/cddis/journal/v7/n3/full/cddis2015409a.html OR https://www.researchgate.net/publication/296698867_Dengue-induced_autophagy_virus_replication_and_protection_from_cell_death_require_ER_stress_PERK_pathway_activation).
If it is toxic try titrating it. Best of luck!
-ED
Article Dengue-induced autophagy, virus replication and protection f...
Hi. Since you have a good delivery system to the targeted cells you might try compound that inhibit electron transport complex Sodium cyanide that coud work for your system. Good luck
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