[Histonet] Double labeling with antibodies that need different fixatives

[Adam .]

I have no idea if it would work with regular formalin. In my experience,
many antigens work as well in zinc buffered formalin without any antigen
retrieval as regular formalin with antigen retrieval. But really, you just
have to try it yourself.

On antibodies I've gotten this to work, I use commercially available zinc
buffered formalin which comes in gallon jugs for around $50. We don't do any
special processing. We plop our sample (bones) in zinc formalin overnight at
4C, decalcify in EDTA or formic acid (only necessary for bones), and then
embed just like any other sample.

Adam

On Mon, Feb 22, 2010 at 9:45 PM, Phebe Verbrugghe <
pverbrugghe <@t> meddent.uwa.edu.au> wrote:

>  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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>
>