If drug inducing ROS or lipid ROS is treated dose-dependently in specific cells, level of ROS measurement by FACS increased dose-dependently?
one thing to keep in mind is that excessive oxidative stress lead to cell death which might affect your results; once ROS levels reach/pass a certain threshold, cells are going to die. now if your treatment is mild, and the culture is able to neutralizes it, ROS will accumulate in the first hour or two before going back to basal levels. So before starting any real experiment, you should do a time course assay with your drug using different concentrations to see which is the suitable for your assay. if you want to do FACS, i would recommend using mitosox an h2dcfda. good luck!
Hi, the theory would say yes; but it's necessary to have a stable and reliable readout for oxidative stress/ROS, which is not an easy task !
Thanks for your discussion! I have one wondering. If so, level of inducing ROS increased time-dependently ? (I don't know reading journal for not more than one time.)
Regards,
one thing to keep in mind is that excessive oxidative stress lead to cell death which might affect your results; once ROS levels reach/pass a certain threshold, cells are going to die. now if your treatment is mild, and the culture is able to neutralizes it, ROS will accumulate in the first hour or two before going back to basal levels. So before starting any real experiment, you should do a time course assay with your drug using different concentrations to see which is the suitable for your assay. if you want to do FACS, i would recommend using mitosox an h2dcfda. good luck!
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