Ii am trying to transfect HUVEC, i have no exptience in transfection, i will make transfection for eNOS gene, what is the best protocol for transfection and after that what is the best method to measure expression of my gene, is western blot enough? Can any one help me in those techniques?
Thanks in advence
HUVEC are hard to transfect cells. The traditional expression system will not work. I would suggest to use Lentiviral system. For detection, WB would be enough.
Thanks Muhammad, but we don't have hood for virus work, Do you suggest to another human cell line?
Hello
Use one of these lipofectamine 2000 and electroporation using nucleofector from lonza.
see the links for protocols steps .
Good Luck
http://www.lonzabio.jp/catalog/pdf/ri/I458.pdf
http://cardiovascres.oxfordjournals.org/content/35/3/522
http://www.molecularinfo.com/MTM/K/K1/K1-1/K1-1-12.pdf
https://www.researchgate.net/post/What_is_the_best_way_to_transfect_HUVECs
http://www.nature.com/protocolexchange/protocols/2096
The Targefect-HUVEC reagetn works well for transfectign all types of primary endothelial cells including HUVECs. You can view details at the following link
https://www.nature.com/protocolexchange/protocols/2096
- Badges
- Science method