Lipofectamine Transfection of BAECs - (Jul/26/2006 )
I am trying to look at apoptosis in BAECs, but before I can do that I need to figure how to get my transfection efficiency up. The best I have gotten is 20% and I have read that I can expect a transfection effciency of about 70%. Does anyone have any tips of how to get better tranfection effciency?
M9 peptide can help increase the transfection rate by facilitaing nuclear translocation; inactivated adenovirus can increase transfection through endosome release.
IF u r susing lipofectamine for transfection, I suggest to transfect the cells immediately after trypsinising them. I am tranfecting my cells (293FTs) in this way and get nearly 95% transfected.
I suspect that M9 peptide or its derivative is the "Plus" reagent sold by Invirogen, but I am not 100% sure.
For trying to get a better efficiency: try different ratio's of lipofectamine vs DNA, try different reagents (if you have time/money). Try diluting your lipofecatmine and DNA in Opti-MEM (if you're not doing that already). Use the Plus reagent, might help. Try to transfect directly after trypsinising as suggested.