Re: GFP / Background Information

<< Previous Message | Next Message >>
From:"tylee" <tylee@itis.com>
To:"Gayle Callis" <uvsgc@msu.oscs.montana.edu>, <histonet@Pathology.swmed.edu>
Reply-To:
Date:Wed, 28 Apr 1999 20:42:47 -0500
Content-Type:text/plain; charset="iso-8859-1"

Histonet,

FYI. There is some background technical information on GFP at...
http://www.clontech.com/clontech/Manuals/GFP/Properties.html.

I believe Clontech now has some sort of exclusive license for use of GFP as
a reporter from Aurora (originally the technology was from one of the UC
labs, but now some of  the scientists [?? Roger Tsien ?? spelling??]are
associated with Aurora).  I am by no means an expert on GFP, but I think I
recall that there are mutant GFPs that are much "brighter" and there are
forms that emit at different wavelengths, like "Blue FP" (that may help
avoid some of the background fluorescence in cells that was near the wt GFP
wavelength).  It is certainly conceivable that some forms could be more
resistant to aldehyde fixation. I recommend a call to Clontech, they may
know the answers.
Ty Lee

-----Original Message-----
From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
To: histonet@Pathology.swmed.edu <histonet@Pathology.swmed.edu>
Date: Wednesday, April 28, 1999 4:40 PM
Subject: more on GFP


>I see the comment "GFP staining".  GFP or green fluorescent protein is
>not a stain, it is a protein, when inserted into a cell, etc, that
>fluoresces with specific wavelength of UV light.  You can have the protein
>in cells, fluorescing near or around the FITC excitation, but is best
>viewed with GFP filters on the UV microscope.  It is visible with FITC
>filters, but we found it much improved with the GFP filters.
>
>You can do double fluorescence work, with GFP fluorescing on its own, then
>perform immunoSTAINING on cells with a fluorescent label (in contrast to
>the Green fluorescent protein) such as TRITC or Alexa 546 (Molecular
Probes).
>
>An example (from a publication on file) GFP in cells, and IFA -
>(immunofluorescent antibody staining) for GFAP glial fibrillary acid
protein
>to see live astrocytes in transgenic mice done on 40 um vibrotome section.
>Very elegant!
>
>Sorry to hog the day!
>
>Gayle Callis
>
>




<< Previous Message | Next Message >>