I have a really old construct that contains GFP (original) tagged protein under CMV promoter. I can't find an empty vector containing GFP (Addgene or similar). The closest they have is eGFP under CMV promoter. Is it okay to use the latter as an empty control. I am not planning to quantify the intensity of green channel, green signal is only used to show transfected cells, so I am assuming the difference in the excitation/emission of GFP and eGFP is not going to affect my experiments. My only concern is the difference in cytotoxicity. Thanks.
Hi,
My experience is that an unrelated protein fused to GFP is a better control that GFP or eGFP alone. It generally induces far less toxicity, probably because GFP alone accumulates in the nucleus and is expressed at very high level.
HI,
I would agree with Camille and would rather use something for the fusion as this will use cell ressources for Transcription and translation in a comparable manner as the other system
Sven
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