[Histonet] Double labeling with antibodies that need different
fixatives
Phebe Verbrugghe
pverbrugghe <@t> meddent.uwa.edu.au
Mon Feb 22 21:45:48 CST 2010
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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