Im doing a I/R model with C2C12 cells and I need to measure ROS production. First I will induce ischemia with the ischemia chamber and then I will measure ROS after 4,6, 8 and 12 hours. In the same timepoints I will induce reperfusion, but I dont know after how many hours after reperfusion its recomendable to measure ROS.
I assume they will be cells in a culture in an aqueous medium where the ischemia-reperfusion experiments will be performed. Therefore I think it would be advisable to start very early after reperfusion. Perhaps a time 0 (baseline before initiating ischemia, then 10 min and 30 min and 60 min after initiation of ischemia, and then 3 min, 10 min and 30 min after reperfusion.
Taking samples of culture liquid or cell suspension solution. Or, directly from a homogenized cells C2C12 subjected to I / R
Thank you so much !
I have done a trial, and yes, for example after 4 hours reperfusion the ROS production decreased. Now I wil repeat it, measuring ROS production 15, 30, 45 and 60 minutes after reperfusion.
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