We have vectors with constructs that are successfully transfected in CHO/Cos cells using the LTX Plus reagent (Life Technologies). Transfecting the same vectors in A431 cells has shown us almost no transfection, even with the GFP control. We want to utilize these cells to study signalling pathways and require a sufficiently high transfection efficiency. Any advice on this would be appreciated.
Yep, here's a transfection reagent specifically for A431 cells that's been optimized for their transfection (https://altogen.com/product/a431-transfection-reagent-epidermoid-carcinoma/). It's based on a lipofection approach that should ensure your vector gets transfected. Be careful with the medium you are using - different cell types can require different conditions for successful growth. If you're buying cells from a company, they should have given you culturing information, and that's your best bet before transfecting the cells at subconfluency.
hi
Its true that A431 is hard to transfect cell line as compared to CHO and Cos. i will suggest to transfect A431 cell using mirus TransIT®-2020 Transfection Reagent. Definitely, you will get good transfection efficiency.
good luck
Try to use TurboFect Transfection Reagent, Its working very well
http://www.thermoscientificbio.com/transfection/turbofect-transfection-reagent/
Best,
Thiru
Thank you Ravindra, Amar and Thirupugal! I have asked for samples from all of the suggested companies to test transfection in our cells.
Has anyone used Fugene 6/HD for transfecting A431s?
we have managed to transfect A431 cells using Nucleofector Kits, a combination of electroporation and chemistry from Lonza. in comparison to the other 2 systems we were using for different mouse and human cell lines th enuclofector system seemed the most efficient. in the A431 cell we managed 30% transfection efficiency with only 5-10% with our other methods. At the time we used the system there was only a general kit for human lines and one for mouse lines. Over the years the have developed different kits and parameters for different cell types. You can check if they have on for the cells you are interested. The only negative is that you need to have the electroporator for this and it can be a bit expensive depending on how many transfections you do routinely. However, it may balance the cost with the efficiency in comparison to other methods.
hope that helps
Yep, here's a transfection reagent specifically for A431 cells that's been optimized for their transfection (https://altogen.com/product/a431-transfection-reagent-epidermoid-carcinoma/). It's based on a lipofection approach that should ensure your vector gets transfected. Be careful with the medium you are using - different cell types can require different conditions for successful growth. If you're buying cells from a company, they should have given you culturing information, and that's your best bet before transfecting the cells at subconfluency.
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