Dear all, I have been struggling with transfection in Hs578t with my pure plasmid ( maxi prep plasmid). I transfected in a 24 well plate , changed the media after 6 hours post transfection. I observe at least 80% cell death within 24 hours. I am wondering whether the Lipofectamine 3000 is very toxic to cells e en with the recommended dose. I used 1.5 ul Lipofectamine 3000 in 25ul. Do you think I should use .75 ul instead? Which one is more toxic between Lipofectamine 3000 and p3000. In my modicum of knowledge, p3000 acts as an enhancer . Has anyone have experience with transfection in Hs578t cells ?
Dear Abir,
I think you used the right protocol, and yes p3000 is an enhancer that you may add to the transfection mix. No importance what is really toxic. But if you observe toxicity, do not go up with the amount of reagent, rather try to use less.
I have no experience with Hs578t and despite my activity of tech support for the Viromer transfection reagents, I have never really heard about these cells. So I do not know if they are difficult to transfect or sensitive to cationic lipid-based reagents as Lipofectamines.
FYI, with our Viromer reagents (polymer-based), we have quite good feedbacks for other breast cells and some published data. See attached file.
Best, Sandra
- Badges
- Science method