transient transfection of GFP fusion

D. Fesquet fesquet at crbm.cnrs-mop.fr
Tue Nov 27 08:17:27 EST 2001


Hi,

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!

cheers

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Dr Didier FESQUET

Centre de  Recherche de Biochimie Macromoléculaire
CNRS UPR 1086
1919, route de mende
34293 Montpellier cedex 5 (FRANCE)
Tel: (33) (0)4-67-61-33-79 ou 33-72
Tel(perso) (0) 4-67-55-67-72
FAX: (33) (0)4-67-52-15-59
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email: fesquet at crbm.cnrs-mop.fr

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