Q:

I have a GFP construct that I use and usually get anywhere from 50-80% GFP positive cells using FACS analysis while I get only 30-40% using a GFP-NFAT fusion protein driven by the same promoter. Do you have any suggestions?

A:

The Amaxa™ Nucleofection™ Conditions are optimised for each cell type, the solutions and programs are cell type dependent and not vector dependant. The expression of the fusion protein depends on the localization of the protein transcription, translation, as well as the folding and stability of the fusion protein. It may also be advisable to change the terminus to which the tag is fused. It is also possible that other Nucleofector™ Programs may help as well. Please contact Scientific Support for further suggestions.

Categories:
Transfection