Hello!
I want to stress osteoprogenitor cells and thereafter measure it, either in the cells or in the media.
Whats the best approach for this?
The best approach i have right now is to use H202 to stress the cells and later use SA-beta gal assay to measure it, but I am unsure if thats the best way.
Edit: I have now concluded that the best measure is to stress the cells using H202 and therafter detect the ROS both extracellular and intracellular using Isoluminol- and luminol enhanced chemiluminescence in cell culture. I have trouble to find a good protocol describing this, anyone familiar with this?
Any input would be valuable, thanks!
Hi Elina,
There are different methods for inducing stress in the cells. Best method would be defined based on your aims. It means that based on the targets that you want to look at after stress induction, you may select specific method. For example, some are interested to study the stress based on unfolded protein response in the cells. therefore they stress the cells either by starving (physiologic methods) or by pharmacologic methods such as Thapsigargin stress induction (there are more options, though). Some are interested to study the stress through hypoxia and the culture the cells in hypoxic environment. If you could elaborate more on your question that you want to find an answer for, you may receive better pieces of advice.
Hope it helps.
Regards,
Zahra
Zahra Sepehri Thank you for your input! Do you have any experience in Isoluminol- and luminol enhanced chemiluminescence for detection of ROS?
I am afraid I do not. I have some experience with starvation and thapsigargin.
Regards,
Zahra
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