RE: Anti-GFP staining in paraffin sections

From:Cynthia Favara


Would you mind letting us know what antibody you use etc?

Cynthia Favara

-----Original Message-----
From: Luis Chiriboga []
Sent: Thursday, May 16, 2002 7:17 AM
To: nina leek; Cathy Gorrie;
Subject: RE: Anti-GFP staining in paraffin sections

Hi Nina & Cathy
There are many reasons that you would want to do GFP staining in FFPE
tissues.  First, FFPE provides superior morphology compared to frozen
sections  and eliminates the need for looking at GFP in fresh material.
Second,  you remove the necessity of having to use a fluorescence scope.
Third, It easier to co-localize the GFP with morphology, and other markers.
We have found that the sensitivity IHC-GFP is comparable to direct GFP
fluorescence detection.
Probably the single  most important benefit (from my experience), is that it
gives you a good idea of what your transfection efficiency is. Plus, in
experimental heterotransplant tumor models you can observe the increase in
heterogeneity (or not) of your implanted cells over time.

Hope this helps!
regards Luis

-----Original Message-----
From: nina leek []
Sent: Wednesday, May 15, 2002 8:35 PM
To: Cathy Gorrie;
Subject: Re: Anti-GFP staining in paraffin sections


It's Green Fluorescent Protein, originally found in jellyfish, and now,
through genetic engineering, available in other colors as well.  Also
genetic engineering, it can be introduced into the genomes of other species,
accompanied by genes for other proteins of interest.  This means you can
screen the organisms you are playing with for the "other protein of
by looking for the fluorescence of the GFP, which is now (genetically) tied
to it.  A very neat trick.

I'm not sure why someone would want to stain for it in (presumably) FFPE
sections, but they may be expansive enough to tell us.

Adrian Leek.

Cathy Gorrie wrote:

> What's GFP?
> At 2:30 PM -0400 15/5/02, Sarka Lhotak wrote:
> >Hello Netters,
> >       Has anybody done a peroxidase staining against GFP in paraffin
> >tissue sections? I am wondering whether the antibody can bind even to GFP
> >that is no longer fluorescent (due to alcohol processing etc.)?
> >       Thanks a lot,
> >
> >Sarka Lhotak
> >
> >Hamilton Regional Cancer Centre
> >McMaster University
> >Hamilton, Ontario, Canada

<< Previous Message | Next Message >>