starvation medium recipe for Hela cells - (Feb/20/2006 )

Some references used EBSS and others used HBSS supplemented with other reagents as starvation medium. What do you suggest for Hela cells? I'm planning to look at subcellular localization of my genes under different conditions. When should I replace the rich media with the starvation medium (before transfection or right after transfection?) and for how long? I want to test both amino acid-depletion and serum-depletion conditions.

thanks for the help.

Hi,
could you please explain me what would be the purpose of determining the subcellular localization of a particular protein. What information can be drawn about the protein and what are the benefits overall?
thanks-

Did you mean you want to see the intracellular trafficking of DNA delivered by transfection reagent under control or starvation condition? If you want to examine cell cycle effect you need to do it before hand, because that takes time for cells to arrest at certain stage. For AA depeletion, that could come faster than cell cycle arrest, you can probably do it after transfection.

You need to have all the controls, because I could see some very messy experiments ahead of you.