Electron microscopy using GUS antibodies

2020001 at SAPHIR.ULAVAL.CA 2020001 at SAPHIR.ULAVAL.CA
Thu Dec 3 16:42:41 EST 1992


Dear networkers

We are currently working with the GUS gene fusion system from Clonetech.
We are studying different promoters by fusing them with the GUS gene and
introducing them into tobacco.  Since our promoter target their gene product
in the embryos (more precisely in the protein bodies) we are also interes-
ted in localizing the GUS gene in the transgenic tobacco plants.  To do so,
we are using the GUS rabbit antibody included in the Kit and we want to
use electron microscopy.  We read in the literature that this kind of
experiment give rise to a lot of unspecific background signal caused by
the binding of the GUS antibody with the lectin proteins present in the
membranes.  The lectin seem to recognize particular sugar on the Gus protein? 
Since we can not eliminate this unspecific binding (we can not purify the 
antibody) does anyone has a solution to our problem?  Have you ever tried
to localize your GUS protein using the antibody include inthe Kit?
Is their a monoclonal GUs antibody disponible commercialy or not somewhere?
We also tried immunofluorescence but we have the same unspecific binding pro-
blem.

Thank you 

Pierre Bilodeau
Dept. of biochemistry
faculty of Sciences
Laval University
Ste-Foy, Quebec, Canada
E. mail: 2020001 at lavalvx1.bitnet



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