Hello, I am trying to induce oxidative damage in neurons (derived from IPSC) by using hydrogen peroxide.After treatment I want to see if there is any evidence of lipid per oxidation (by looking for malondialdedye) or if there is any evidence of 8oxoguanine accumulation. can anyone advised how long I should wait after treatment before assessing for these?
thank you
I wonder how you would determine accumulation of MDA ex vivo while you may not have info regarding baseline/level of total antioxidant (TA) present, it is virtually indeterminable!
Be reminded that;
1. MDA development is a function of how much TA already present. This could only be determined by taking baseline, I wonder how you could this in animal models with limited blood supply.
2. MDA begins to increase immediately you induce with radical-generating substances, only curve may inform you proper how fast this takes place, consequent upon TA concentration in the body which is a function how sufficient nutrients had been earlier been uptaken to the system.
Finally, I advise;
1. Search for literature to see previous procedure in doing this.
2. Or set a timeline after overloading with peroxide, and report it so-so hourly e.g injected 4-hourly for 4 days, depending on your topic. Be informed that it takes much time before reactive oxygen species from lipid peroxidation do substantial damage to the DNA and initiate cancer development.
SUMMARY: IT DEPENDS ON HOW MUCH TOTAL ANTIOXIDANTS INITIALLY PRESENT, WHICH MAY BE HARD TO KNOW EX VIVO WITHOUT BASELINE ESTIMATION. BASELINE ESTIMATION MAY BE BIT DIFFICULT IN ANIMAL MODELS WITH LIMITED BLOOD SUPPLY.
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