[Histonet] Double labeling with antibodies that need different fixatives

[Phebe Verbrugghe]

Hello Adam,
 
Thank you very much for this very useful information!
Do you know whether this would also work on tissue fixed with formalin
instead of zinc buffered formalin by any chance? 
Also, could you give me the recipe for the zinc formalin and can I use a
standard tissue processor for embedding in paraffin or should I use a
specific protocol manually and if so, which? 
 
Thanks!
 
Phebe

________________________________

From: Adam . [mailto:anonwums1 <@t> gmail.com] 
Sent: Tuesday, 23 February 2010 10:52 AM
To: Phebe Verbrugghe
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Double labeling with antibodies that need
different fixatives


Although I haven't tried it myself, others have gotten CD31 from BD to
work on FFPE tissue using the tyramide amplification system on zinc
buffered formalin fixed sections. I've generally had good luck with zinc
buffered formalin myself for many antigens so it may work for your other
one.

See http://www.ncbi.nlm.nih.gov/pubmed/19052548

Just to be clear, they used zinc buffered formalin, which isn't the same
thing as zinc fixative.

Adam


On Mon, Feb 22, 2010 at 8:41 PM, Phebe Verbrugghe
<pverbrugghe <@t> meddent.uwa.edu.au> wrote:



	Hi all,
	
	I would like to do an immunofluorescent double labeling with two
	antibodies but 1 antibody works on acetone fixed frozen tissue
but not
	on formalin fixed paraffin embedded tissue (CD31 BD pharmingen
553370)
	and the other one works on formalin fixed paraffin embedded
tissue but
	not on acetone fixed frozen tissue. Is there any way I could
still do a
	double labeling and how?
	Also, does anyone have experience with zinc fixative? If my
antibody
	works on formalin fixed tissue is it likely to also work on zinc
fixed
	tissue?
	
	Thank you very much in advance,
	
	Phebe
	
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