I am trying to tranfect CHO cells with the murine FcgammaRIII. I cotransfect it with the murine gamma chain as I have read that this is required for protein expression. I have tried lipofectamine, Fugene and x-tremegene at varying conditions but it does not work. I have tranfected human FcgammaRIIIb receptor in the same CHO cell line without any problem both with -tremegene and lipofectamin. Does anyone have any ideas on why my expression/transfection does not work.
Transfection of CHO cells with Fc receptor
Started by AnetteM, Jun 01 2011 11:44 AM
1 reply to this topic
#1
Posted 01 June 2011 - 11:44 AM
#2
Posted 02 June 2011 - 02:06 AM
Hi,
I too came across a similar situation, in which i could express a contruct in 293 or Hela but couldnt make it express in some of my cancer model cell lines which is definetely not transfection resistant (though not highly competent like 293), as i could express some other plasmids with ease. the only difference i could say between these cell lines is that my cancer models has endogenous expression whereas the 293 or hela doesnt...so my concern is would the cells wont allow to express the exogenous if it already has the same protein highly expressed endogenously???...if anybody came across such a phenomenon and if you have any explanation for this please shed your inputs...thanks in advance...bye..
Gnana...,
I too came across a similar situation, in which i could express a contruct in 293 or Hela but couldnt make it express in some of my cancer model cell lines which is definetely not transfection resistant (though not highly competent like 293), as i could express some other plasmids with ease. the only difference i could say between these cell lines is that my cancer models has endogenous expression whereas the 293 or hela doesnt...so my concern is would the cells wont allow to express the exogenous if it already has the same protein highly expressed endogenously???...if anybody came across such a phenomenon and if you have any explanation for this please shed your inputs...thanks in advance...bye..
Gnana...,
Edited by GNANA, 02 June 2011 - 02:09 AM.
I would prefer being perfectionist rather than a passionist in Research.
I always had an alternate hypothesis....
I always had an alternate hypothesis....













