GFP and immunofluorescence

anonymous abyss at mail.utexas.edu
Thu Apr 17 11:07:10 EST 1997


> 
> I have also made the observation that the GFP fluorescense is lost in cells
> fixed with methanol.  In this case I expressed the GFP directly from an
> unmodified pEGFP-N3 or pEGFP-C1 vector (Clontech).  However, when
> expressing the GFP as a fusion protein (with the GFP at the C-terminal end
> of the construct), I was able to fix the cells with methanol without any
> loss of signal.  I have used about 10 different fusion proteins so far and
> it worked with all of them.  You might try to just clone any protein in
> front of the GFP.
> 
> Hope this helps,
> 
> Gunther


I have been trying to make IMF slides with GFP-fusion cells. It has been a
problem in keeping green signal as it was after fixation with
formaldehyde.
I believe that you must have some sort of idea on how to fix the cells.
May i ask for any help on that???

Thanks



Jae kim
UT at austin
abyss at mail.utexas.edu



More information about the Methods mailing list