I can possibly think of several causes: 1) stability of H2O2 I used, even I prepare fresh solution every time. 2)seeding cell density. 3) culture time between seeding cells and adding H2O2, which is not provided in the reference I followed. 3)incubation time after adding H2O2, I use 3-4 hours.
I do not know where the problem is and what I should do next. Anyone can give me some suggestions?
It seems a mess here. Thanks a lot for help.
I have some experiment details here for both experiment:
cell density: 40,000/well and 15,000/well (both 96 plate)
culture time between seeding cells and adding H2O2: 2 days and 3 days
incubation time after adding H2O2: 3 and 4 hours.