RE: GFP/IHC unclear

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Date:Wed, 13 Oct 1999 10:50:49 -0500
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Neal and others interested in GFP:

We've used the eGFP variant with excellent results. It's fluorescence
withstands alcohol dehydration, clearing with methyl salicylate and paraffin
infiltration. We then cut and stained the sections with eosin as a
counterstain (this process involves more alcohol and organic solvent
exposure) and still got excellent fluorescence. I can send a *.tif file for
you to examine. Further, I have examined slides that I had stained
immunohistochemically using anti-GFP under fluorescence conditions and still
see the GFP fluorescence. I have not tried NaBH4. I'm not sure why such a
strong redox chemical is required for IHC perhaps your milk protein epitopes
are difficult to unmask.I beg forgiveness for my ignorance there. I have not
yet examined the slides I did IS-PCR on for fluorescence. I'll do that soon
and let you know. I looked for the gene insert that the GFP reported. That
worked well but I need to take the slides to the confocal to look for a
green glow. Excellent idea. Thank you. Hope this helps.

Let me know if you want to see the *.tif file with GFP and eosin
fluorescence. It's rather pretty.Good luck,

Donna Montague
UAMS Orthopaedic Research
4301 W. Markham St. Slot 644
Little Rock, AR 72205

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