ATP depletion

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Tiffy
Tiffy's picture
ATP depletion

Hello,
I would like to carry out an ATP depletion with MCF7 cells to investigate a protein translocation into the nucleus. I have found a publication where the authors described a method. But one point I didn`t understand. They used dialyzed serum and I wonder why. Can anybody answer that question? Thanks a lot.
Tiffy
PS It would be very kind if there other advice concerning ATP depletion. What kind of pitfalls etc.

Jason King
Jason King's picture
Dialyzing would reduce the

Dialyzing would reduce the concentration of small molecules such as ATP etc.

samm
samm's picture
Dialyzed serum has much lower

Dialyzed serum has much lower levels of small molecules such as ATP, as well as minerals such as Mg, Mn etc that can affect specific rxns. Most manufacturers (HyClone, Sigma,Gibco etc) sell dialyzed serum in small bottles that you can use specifically for these experiments.
Ensure that you condition your cells beforehand!