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urgent (sorry) : Fixing GFP for phalloidin stainining of cytoske

duerrj at omrf.ouhsc.edu duerrj at omrf.ouhsc.edu
Wed Mar 11 16:01:23 EST 1998


Molecular Probes sells a rabbit anti-GFP polyclonal which works
on acetone and on methanol fixed tissue.  I have found it to be
extremely specific, with a conjugated secondary it is almost as
bright as plain live GFP.

Janet Duerr
Program in Cell and Molecular Biology
Oklahoma Medical Research Foundation
In article <199803111244.MAA13330 at intwww.zit.med.uni-tuebingen.de>, wgschech at med.uni-tuebingen.de ("Wolfgang Schechinger") writes:
> Hi all. 
> 
> Since acetone will kill GFP fluorescence: How will I get the 
> cytoskeleton of 293 fibroblasts and smooth muscle cells stained? 
> Rhodamine-phalloidin and coumarin-phalloidin work well on acetone 
> fixed cells, but in methanol - nope. How might I get out of this 
> dilemma? Would adding the phalloidin dye to the culture medium help, 
> depite it will hurt cells?
> Could I use another fixing procedure, actin dye or what should I do?
> 
> Standing by for your suggestions!
> 
> Wolfgang
> 
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> Wolfgang Schechinger         
> University of Tuebingen
> email: wgschech at med.uni-tuebingen.de
> http://www.medizin.uni-tuebingen.de/~wgschech/research.htm
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