MDA-MB-231 cell transfection - (Jun/01/2011 )
I've tried to transfect MDA-MB-231 with lipofectamine 2000 and it didn't work. I've been using 1ul lipofectamine for 3wells (of a 24plate) because it is the amount my labmates are using with another cell lines to avoid citotoxicity.
I've seen that invitrogen has a special lipofectamine (Lipofectamine™ LTX Reagent) for this kind of cells but it seems that you need to collect cells between 18-24hours after transfection and i would need to collect them after 48hours. Does anybody transfect these cells before and can tell me with reagent/amount has used?
Have you worked with Lipofectamine™ LTX Reagent before and know how it works?
I did Hela, HT1080, SF188 cells. The LTX plus reagent seems better than 2000 in my hands, but I never tried MDA-MB-231 before.
Check out nanoTherics Ltd, they use nanoparticles and oscillating magnetic arrays to enhance transfection efficiencies in a wide variety of cell types.
Heres the webpage for further details: http://www.nanotherics.com/
You really need to titrate the amount and ratio of DNA:transfection reagent for each cell line and each plasmid you use. What works for one will not always work for another.
I have used Lipfectamine, lipofectamine 2000, Lipofectamine LTX, Fugene 6 and PEI for various transfections of DNA and there is always some cytotoxicity. Personally I have seen the greatest cytotoxicity with the lipofectamine variants, and the least with PEI, but results vary between researchers.
thank you all guys! after three times without any result, i'm gonna try electroporation. but thanks, though.
I've been searching for some electroporation protocols, and everyone I've found is for 48 well plates or 6 well plates. I'd like to try 24well plates, due some cells would die during electroporation, i'm not sure about how many cells should i seed per well...
I'm using gene pulser Xcell from biorad in 0.2cm cuvettes, with a concentration at electroporation time of 1x106cell/ml, but no idea about how many cells should I electroporate and seed for each well. any idea?