RE: green fluorescent protein

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From:MontagueDonnaC@exchange.uams.edu
To:Cathie.Crukley@sjhc.london.on.ca
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Cathie: You may be able to do just that. If the green fluorescent protein
used was the EGFP or enhanced gfp from clontech. EGFP is stable for paraffin
processing. We used it as a reporter for a gene we inserted into bone marrow
stem cells. We fixed in formalin, decalcified with 5 % formic acid and
processed on our usual schedule. Cut the blocks at 5 microns, deparaffinized
the slide in three changes of xylene and mounted with Permount and a no 1
coverslip. The fluorescence was crisp and easily located both by standard
fluorescence microscopy and by confocal scanning laser microscopy. Details
and pictures will be presented in my workshop at NSH this fall in Milwaukee.
Hope this helps, Donna Montague, UAMS orthopaedic Research, Little Rock, AR

-----Original Message-----
From: Cathie Crukley [mailto:Cathie.Crukley@sjhc.london.on.ca]
Sent: Tuesday, July 18, 2000 10:18 AM
To: histonet@pathology.swmed.edu
Subject: green fluorescent protein



Hi,

I know nothing about green fluorescent protein, but one of our researchers
has some retina in formalin that has been labelled with a green fluorescent
protein tagged antibody and wants to know what to do with it next.  His
intention is to paraffin process and section it and view it under the
fluorescent scope.  Or will he have to stain it otherwise?

Thanks,
Cathie

SJHC , London, Ontario




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