1 reply to this topic

[burcin]

Hi everybody.
I need help about transfection of insect cell lines. I am not used to them. Cell line is a CNS line (adherent). I use MT promoter. I prepared EGFP control plasmid and transfected, the result was nothing. I also used Ac5 promoter (for insects) with EGFP, the result was nothing again. I tried this Ac5 in 293FT (mammalian) cells and GFP was present (but expression was low). So I thought that the problem was about transfection. I use Turbofect reagent, I will also try other reagents. However this should not be an "all or none" process. I cannot go on transfection of my fused protein.
Is there anyone who can tell an efficient protocol /reagent in adherent insect cell lines?

[protolder]

burcin, on 29 April 2011 - 06:00 AM, said:

Hi everybody.
I need help about transfection of insect cell lines. I am not used to them. Cell line is a CNS line (adherent). I use MT promoter. I prepared EGFP control plasmid and transfected, the result was nothing. I also used Ac5 promoter (for insects) with EGFP, the result was nothing again. I tried this Ac5 in 293FT (mammalian) cells and GFP was present (but expression was low). So I thought that the problem was about transfection. I use Turbofect reagent, I will also try other reagents. However this should not be an "all or none" process. I cannot go on transfection of my fused protein.
Is there anyone who can tell an efficient protocol /reagent in adherent insect cell lines?

Hola, I have worked with the classical cell lines for baculovirus infections, at the begining I used commercial liposomes reagents but for me polietilenimine reagents (Jet-Pei) are easier to use with good results. but in all the cases the tranfection efficience varies, but to have an stable transfected cell line you have to choose recombinant cells adding the selection marker of plasmid. Buena suerte