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Stable cells survive with antibiotics but no exogenous expression detected with - (Dec/21/2011 )

Hello everybody,
I have transfected Hela cells with histagged 45kDa protein in pcDNA3 and selected stable cells with geniticin for 3 weeks. At the end selection, I could not detect any exogenous protein with WB, altough all of negative control cells (no DNA) have died.
What may be the reason? Any suggestions or ideas are appreciated,


Hello. I'm no good in chemistry, but maybe you have some acid in those cells?
Or soap? Maybe you dropped something in there?
Just saying.
mkv player


Hi mWright.
I have tried stable transfection for several times, this is not for the fisrt time, so it can not be accidental I guess. I have transfected 4 different mutants and a wild type of my proteins and with all of them I got no exogenous expession at all :(
I am using Lipo2000 and selecting with 500ug/ml geneticin. After 24 hours of transfection I split my cells into bigger plate, as they become too confluent. But I can clearly observe that -DNA cells are dying but others survive.
Have no idea what is going on....