Medium used after transient transfection?
Posted 01 April 2011 - 11:15 AM
After performing a transient transfection, would it make a difference if I change the transfection medium with reduced serum medium (0.1% FBS) rather than medium with 10% FBS ? Would this affect the expression of my transgene? I would like to incubate my transfected cells in reduced serum, stimulate with my growth factor and then harvest for WB analysis. I'm considering the 24-72h window period, but I don't know if my transgene can still be expressed in reduced serum/serum free conditions.
Any advice/comments would be greatly appreciated.
Posted 02 April 2011 - 11:25 AM
I would rather suggest to express your gene normally, and then 1) use a defined serum without your growth factor, 2) incubate your cells 2 days at normal condition, then make a serum starvation for 4hrs before feeding your growth factor or 3) don't give a damn about the levels of growth factor in the serum, but feed some extra growth factor anyway. That would be what most people do. I routinely do option 2), but I'm into immediate-early response genes...
Edited by Rsm, 02 April 2011 - 11:25 AM.