Stable cell line
Posted 02 November 2012 - 12:19 PM
I am trying to generate a stable MARC 145 cell line. I successfully transfected the cells with phCMV vector containig the gene of interest, transiently. However, after selecting with G418 for about 1 month, the Immuno fluorescence and WB analysis does not show the expression of the protein.
Any of you have an idea what/where the problem could be?
Please let me know your suggestions.
Posted 02 November 2012 - 04:57 PM
How was the initial expression of your protein of interest? And what was the transfection efficiency?
Have you done a kill curve with your untransfected MARC cells? you sure that your current concentration kills all cells (=nothing left in the plate)? This is very important as you don't want untransfected cells to recolonise your culture.
Edited by science noob, 02 November 2012 - 05:00 PM.