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GFP fixing

Steffen Dietzel dietzel at sun0.urz.uni-heidelberg.de
Wed Apr 2 02:58:16 EST 1997


I did not do any quantitative measurements and I should mention that I
work with a very bright GFP-signal. When I tried it I could see the
GFP-Signal very well after 5 min in Methanol. However, morphology of the
specimen is a different thing...
I think I have not tried Ethanol after I found out that buffered
Formaldehyde works fine for the fluorescence signal and quite well for
morphology 


Steffen

In article <333F4E9E.DE2 at sprintmail.com>, Tom Frey
<tomfrey at sprintmail.com> wrote:

> Steffen Dietzel wrote:
> > 
> > (Robert Means) wrote:
> 
> > >         Can anyone give me a pointer to a protocol for visualizing GFP
> > > infixed tissue? Thanks in advance.
> > >
> > > Bob Means
> > 
> > You can use about every fixation method you want to, as long as pH is more
> > or less neutral. Acidic Acid does not work for example. (Buffered)
> > formaldehyd is fine.
> > 
> > Steffen Dietzel
> > 
> 
> Steffen
> 
> I have heard/read other investigators suggest that ethanol fixation
> works very
>  poorly for GFP.  They suggest that the protein is either destroyed or
> not well
>  retained upon rehydration.  Could you let us know if this agrees with
> your
>  experience?
> 
> Tom

-- 
Steffen Dietzel
Zentrum fuer molekulare Biologie Heidelberg (ZMBH)
University of Heidelberg
Im Neuenheimer Feld 282
69120 Heidelberg, Germany
Phone: +49/6221/54-6836 or -6879. Fax: -5893
listservmail:dietzel at sun0.urz.uni-heidelberg.de
personal mail: dietzel at wotan.iwr.uni-heidelberg.de



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