BHK Cell Transfection - (Jan/09/2008 )

I'm trying to optimise transfection of constitutively active G protein cDNAs into BHK cells and use them as a readout for SRE luciferase (stably expressed within the cells). So far I've tried fugene 6 but either the transfection isn't working or I'm missing the peak of the SRE response. How long post transfection would you lyse and read the luciferase values bearing in mind the cells will not be ligand treated? Would you serum starve the cells at the time of transfection? Also, has anybody optimised a fugene 6 transfection method for this cell type?

Thanks in advance.

Colin.

24-36 hrs post transfection. Since you are using a derivative line of BHK, you should optimize it yourself.

we checked luciferase activity after 48 hrs. we didnt really starve cells at time of transfection. We had them in 2% serum.