RE: GFP

From:Emma Carter <E.Carter@oxfordbiomedica.co.uk>

Dear Angeline,

You may well find that if you paraffin embed you lose the fluorescence,
although you might be able to use antibodies to detect it (which of
course, opens up more problems!). we have used both fluorescence and
antibodies to detect this protein, and have managed to accomodate most
problems, (although if anyone can suggest ways to prevent non specific
staining of GFP in muscle, like other antibdies, i would be grateful!).

You should be able to see it with UV in the frozens, we certainly have
with GFP in muscle and in tumours, though we havent really looked so
much in brains...

Let me know if i can help any more,

Emma Carter
Oxford BioMedica (UK) Ltd

> -----Original Message-----
> From:	Angeline Martin-Studdard [SMTP:AMARTIN@mail.mcg.edu]
> Sent:	Tuesday, January 09, 2001 1:20 PM
> To:	histonet@pathology.swmed.edu
> Subject:	GFP
> 
> Hello All,
> 
> I will soon be delving into the visualization of GFP positive cells
> that have been transplanted into normal mice.  I have seen a few blips
> on the histonet regarding problems with this protein.  I will be
> working with brain tissue sections and will try both frozen and
> paraffin embedded.  I'd like to know if anyone else has experience
> with this.  Can the fluorenscence from the protein still be seen after
> processing of the tissue or does it require detection and/or
> amplification? 
> 
> Any advice or assistance you can offer will be very helpful.
> 
> thank you,
> angeline
> 
> angeline martin
> cell biology and anatomy 
> medical college of georgia
> augusta, ga
> amartin@mail.mcg.edu



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