I'm using G418 to abrogate translation in 293 T cells completely and as soon as possible (I'm not doing selection). Concentration that I use is 3,5mg/ml. However I'm not sure that translation is fully blocked even after 6 hours of incubation. I would greatly appreciate your comments and experience about how fast and at what concentration G418 inhibits protein synthesis entirely.
As far as I know, G418 is pretty slow and I do not have a strong expertise with it. But since it is not for selecting resistant cells, I will recommend you use puromycin or cycloheximide, which are a way more rapid and efficicient. Good luck
Yikes! 293T are already G418 resistant (T is for T antigen, which was selected with a G418 cassette).
Oh I see I didn't know that and obviously it is the reason, I will look for HEK293, thanks.
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