free style 293 cells - (Sep/08/2005 )
Dear all,
because i am going to purify protein complex at later stage. so someone recommended me to use free style 293 cells, invitrogen rather than 293T cells which normally grow monolayer. but this free style 293 cells grow suspension, so can get the cells lots. but i do not know whether freestyle 293 can be stably transfected with my gene, but I do know transfection reagent is good for making stable 293T cells.
Anyone give me some suggestions, anyoneof you did stable transfection on frees 293 or ever used it?
Thanks a lot
FreeStyle 293 system is used for large scale transient transfection and protein expression in suspension 293 cells.
Stable transfection is also okay, but not recommended because this system is not specially used for this purpose.
Please consider whether you would like to have protein expression easily and quickly OR stable expression.
What is the purpose for stable expression?
You would like to have further research?
If you simply would like to have large scale protein expression within a short period of time, FreeStyle 293 would be a great choice.
If you would like to have STABLE transfection for further analysis, you are better to look for another system.
because i am going to purify protein complex at later stage. so someone recommended me to use free style 293 cells, invitrogen rather than 293T cells which normally grow monolayer. but this free style 293 cells grow suspension, so can get the cells lots. but i do not know whether freestyle 293 can be stably transfected with my gene, but I do know transfection reagent is good for making stable 293T cells.
Anyone give me some suggestions, anyoneof you did stable transfection on frees 293 or ever used it?
Thanks a lot
I am going to purify protein complex which going to use lots cells. for 293T cell line as a monolayer , I probably need to grow lots of flasks and so on. that is the reason freestyle 293 cells has the advantage. at the same time I want a stable transfection with an inducible system other than transient transfection, so i can control the induction. do you think at this circumstance I better not use freestyle 293?
Thanks a lot
Thanks a lot
As said before, 293 Free ar NOT meant for stable transfetion, they are only for quick, small to medium scale manufacture of proteins. If you just want some protein they are a LOT faster than making stables and then harvesting from adherant 293 cells.
If you want a stable inducible system then you have to go with regular 293 or 293T. If you use the 293T, however, remember that they already carry a resistance marker (neo I believe, but look it up) that was used to put in the large T antigen so make sure you use a different selectable marker.
you are right. there is a neo in normal 293 T cell lines. I did not understand what you said abut large T antigen. I think I can use it to select the stable transfected cell lines using geneticin. what is to do with that large T antigen? probably a stupid question, but I have no idea what is it.