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Position: Discussion forums > Bioscience biotechniques discussion forums > Cell biology and histology methods forum
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ATP depletion
Posted by: Tiffy (IP Hidden, New member, 4)
Date: February 26, 2008 12:18PM
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.
Re: ATP depletion
Posted by: aphio-igor (IP Hidden, New member, 6)
Date: March 16, 2008 05:03PM
I'm in no way claiming more than a passing interest in the ATP end of things, but since I'm new to this forum, I thought I'd offer an educated guess for why dialyzed serum:
Serum is chock full of growth- promoting 'goodies' that, for normal in vitro application, is great. The problem exists when doing metabolism/ nutritional studies, or studies using labeled material that may also be in the serum. As serum is a natural product, the relative amount(s) of 'goodies' are likely off from one bottle (or at least lot) to the next. To cut down on variability - as well as diluting out the incorporation of whatever labeled component they're studying - the whole serum is dialyzed to remove the component in question. I hope that made sense for you. Best wishes to you and this project.
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