I would really appreciate your opinion about this problem I've faced.
I cloned into PcDNA 3.1 Hygromycin and Zeomycin. The insert sequence as well as the promoter regions were sequenced and they're perfect!
I transfect these vector into Mouse embrionic Fibroblast using Lipofectamine LTX that is quite good for transfect sensitive cells. The transfection rate was stablished at 1:7
Five days after transfection we start the selection.
I ran PCR and the cells were positive as well as they're growing pretty fine under the presence of the highest antibiotic concentration.
On this way that should be expected that these cells would express high levels of my protein but when we screen the cells by FACS they are all negative.
I haven't tested in Western Blotting but anyway they should be positive in FACS.
I would really like your comments for this problem
Edited by Cell buster, 21 October 2013 - 04:07 AM.