Effect of radiation in hypoxic and acidic cells - how to initiate the experiment about protein expression in hypoxic and acidic ca (Mar/03/2006 )

I want to see the effect of radiation on the hypoxic and acidic pH growing cells. so what should I do??? after cell seeding ,next day first change the medium to low pH media and irradiation and put in the hypoxia chamber or anything else??? How can i make the control?? I want to see the protein expression in different time intervals.I am in confusion,please help me.

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hi, i think it depends on what you are going to test. As you stated, i think it is better to do hypoxia and low pH growing first and then do radiation second. Well of course it is even better if you have the equipment to do both of radiation and hypoxia at the same time. Just personal opinion, good luck! How long are you going to expose your cells under radiation?

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HI Superpk80
thanx a lot for your interest.
Because tumor cells are hypoxic as well as growing in acidic microenv in vivo condition but i am doing this experiment in vitro so first I want to make my normal cells hypoxic and acidic by incubation in hypoxia incubator in low pH medium. so that the conditions in the experiments shud be at least comparable to in vivo. What i want to kno is that to make the cells hypoxic how long shud i incubate the cells after changing the normal pH media to low pH media???so that the cells behave as hypoxic and low pH adapted cells. I wanna detect the protein expression induced by radiation in the hypoxic and low pH cells. I hope everything is clear now.I have the radiation n hypoxia chamber separate so I can not do at the same time.I am using 4 Gy of radiation dose for 48 secs.
What are your valuable suggestions for me??? please feel free to write.

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