colocalisation

Hans Peter Lauritzen H.P.Lauritz at mfi.ku.dk
Mon Oct 11 08:05:33 EST 1999


Katryn Sunn wrote:
To: fluorpro at net.bio.net
From: k.sunn at garvan.unsw.edu.au (Kathryn Sunn)
Subject: COLOCALISATION WITH GFP
Date: 10 Oct 1999 22:52:34 -0700
Dear All


Has anyone done a lot of colocalisation studies using GFP tagged
transfected constructs and a red-tagged antibody? If so has anyone ever had
the problem of the GFP green signal losing its fluorescence after
permeabilisation? Does anyone know any handy hints to get around this at
all?


Thanks for your help


Kate


___________________________________________________________________
Kathryn Sunn BSc (Hons)
Postgraduate Student
Bone and Mineral Research Program
Garvan Institute of Medical Research
384 Victoria St, Darlinghurst
St Vincent's Hospital Tel: (02) 9295 8260
Sydney NSW 2010 Fax: (02) 9295 8241
Australia email: k.sunn at garvan.unsw.edu.au
___________________________________________________________________



----------------------------------------------------------------------------
----

Hi Katryn
It proabely depends on your procedure. Whitch procedure are you using?
I have immunostained the transferrin receptor with texas-red in GFP-fusion
expressing primary muscle cells, with out any problems with the EGFP
fluorescence. I use fist 2% paraformaldehyde to fix followed with saponin
for permeabilisation and finally the antibodies with saponin. I mount in
vectashield. I have not seen any decrease in EGFP fluorescence on specimen
even after 1½ year in the fridge. 
		Med venlig hilsen 	Hans


Hans P.M.M.Lauritzen, B.Sc., M.Sc., Phd-student.
Copenhagen Muscle Research Center, The Panum Institute 12.4 
Copenhagen, Denmark
E-mail: H.P.Lauritz at mfi.ku.dk
Phone: +45 35327375 or +45 35327436 or +45 35327430
Fax: +45 35327420



More information about the Fluorpro mailing list