Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Cloning

Is pFLAG expression toxic? - FLAG seems to kill my HeLa cells. This can't be right (Mar/29/2010 )

I cloned my gene of interest into pFLAG and then transfected into my HeLa cells. Expression of this gene normally leads to apoptosis. Surprisingly I am also seeing cell death in control wells which only have the original vector itself with no insert.

This seems odd because in wells that I have Lipofectamine without any vector I don't see any cell death. So it is like pFLAG expression is causing cell death.

I used 1-5 ug of DNA for my 6-well plates.

Does anybody have experience with FLAG?


Something toxic in your plasmid prep? Have you tried with a different vector such as puC?