I am working on the patch-clamp recording now. But I cannot find a appropriate amount of Lipofectamine3000 to add into the cell when transfection. I can't use the normal proportion that suggested. The lipo is harmful to the cell membrane. I need some advice.
Hello! Could you give us some more information to help you?
What are your cells: neurons, muscle cells, pancreatic cells?
Do you transfect siRNA or DNA?
It could be possible that the lipofectamine is toxic to your cells because it's a lipid-based reagent. Then interfering with your cell physiology.
A great alternative would be to work with polymer-based reagent instead.
If you want to test our Viromers for example, let me know, I can provide you a free test sample.
Waiting for more details, please.
Bests
ok, thanks for the info.
HEK cells are very easy to transfect, whatever the reagent you use.
If you observe toxicity, all you can play on is the reagent/DNA ratio to try using as less reagent as possible.
I maintain that your problem is probably linked to the use of a lipid-based reagent then interfering with your cell membrane. I do not know exactly which modifications are induced by patch-clamping, but there is something with the cell membrane for sure.
Did you try first to transfect the cells not under patch-clamp recording?
Again, if you want to test our Viromer RED (polymer-based reagent), please tell me, we have a distributor in Tapei, they can provide you a test sample very rapidly. This reagent has very high efficiency on HEK cells (>80%) with no sign of toxicity.
And to be honest, I will be more than curious to see how it works in such experiments!
Bests, Sandra
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