transient transfection of GFP fusion

D. Fesquet fesquet at
Tue Nov 27 08:17:27 EST 2001


I am studying a protein localized at the centrosome by indirect IF
with specific
Ab. in order to do a structure/function study, I want to use GFP
fusion. unfortunatly, after transient transfection, The only result
is a green cell floaded with a GFP signal, probably due to the
overexpressed protein...
so there is no hope I can do time lapse imaging of these cells.
does anybody manage to get rid of the proteine made in excess in the
cell.  I am told, that first extracting then fixing the cell....can
get rid of the unspecific staining. anyone experience with this?
any help is wellcome!



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