Stable transfection using pCMV-FLAG2 - has anybody done this? (Aug/22/2006 )
Hello all, I was wondering if it is posible to stably transfect a mammalian cell line (MCF-10A) with pCMV-FLAG2-protein. I have no experience in stably tranfection, so i dont know if it matters the kind of vector is used (I suppose it does). I need the flag tag for protein purification and immunoprecipitation studies.
What do you think?
i think you can do a selection with this vector as there is the Neomycine resistance gene carried by this vector. The limits are the b if your protein is toxic, or growth inhibiting. These points will drive a hard selection procedure.
I already used pCMV-FLAG vectors for stable transfections and they really worked fine for my proteins...
Good luck - after all you are going to spend quite much time at the flow now...
Thanks for the comments, thats good to know. My protein is not toxic, so, I dont see any problem then.