Lipofectamine LTX

[darkwng]

Hi,
has anyone tried the transfection reagent Lipofectamine LTX from Invitrogen?

Invitrogen claims its better than FUGENE. I use Fugene so I was wondering if it is in fact better claim in terms of induced cytotoxicity and percentage of positive transfections?

thanx

[herry]

Lipo LTX is definitely better than Fugene 6 and HD for most of cell lines.

[MikeHolloway]

Definite cytotoxicity with Hela using Plus with the LTX. Invitrogen protocol calls for leaving Plus and LTX mix on the cells. The resulting cytotoxicity isn't enough to kill the cells instantly, plasmids express their proteins, but they definitely stop dividing, and some enter what looks like apoptosis in time lapse image capture at about 48 hours. Don't know yet if limited exposure works with a decrease in cytotoxicity. Trying about 3 hour exposure.

[xangwang]

Same problems of toxicity. can be avoided by reducing LTX quantities and adding combimag (ozbiosciences). magnetofection allow to reduce lipid doses while raising the efficiency

w.

[rp0497]

Hi xangwang. How does magnetofection improve LTX efficiency ? do you have any reference about it, or have you experienced it yourself ?

thanks in advance

RP.

[xangwang]

Hi rp !

Don't know exactly how that work but from what I've read, LTX (or L2000 or any lipid reagent) and nucleic acids forms lipoplex. Then that lipoplex combines with the magnetic nanoparticle (Combimag).
Under your cell plate you put a strong magnetic plate that attract the magnetic particles linked to lipoplex.
The point is, 30 min of incubation are suffisant to reach a high transfection efficiency, so you can perform a medium change very earlier, compared to the 6h needed with LTX use. So the toxicity is reduced while efficiency is amplified.
I've tested Combimag in association with either L2000 and LTX and it works very well .
pv me if you need more info

w.

[rp0497]

thanks for the tip, it sounds very good indeed if you can reduce toxicity and improve transfection at the same time ! I will definitely try and let you know about the results.

Best

rp.