What are the best GFP tags?

Fraser D. Brown F.D.Brown at bham.ac.uk
Wed Nov 13 13:47:10 EST 1996


Dear all,

I will shortly be tagging two proteins with GFP. Both are mammalian
phospholipases which I want to transfect into cells together and
separately and visualise both. What I'd like to know is


1. Which two GFP mutants would you rate highest e.g. S65T, humanized ,
blue-shifted, red-shifted , etc

2. Would you recommend N-terminal tagging or C-terminal tagging,

3. Is there a difference in ease or efficieny of transfection between
kits/mutants

4. Taking these into consideration, which kit do you recommend?

and, finally, what would be the best way to perform co-localisation
studies with the GFP tagged proteins and antibodies targeted to other
proteins?

Thanks in advance,

Fraser
F.D.Brown at bham.ac.uk



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