slides and RE: [Histonet] Low temp Antigen Retrieval for Bone Sections

[Gayle Callis]

I strongly suggest if you try this retrieval, that you use Erie Scientific 
EXCEL slides, which are coated specially to reduce the effects of chelator 
based retrieval solutions at high aka alkaline pH.  Many people experience 
section loss with this pH and chelator, so Erie developed a new positively 
charged coating to counteract losing precious sections.   You can access 
samples from them to see if they work out for you, contact your Erie sales 
rep or even Erie directly, they have a website.

Biogenex also has a decalcified bone retrieval solution, another one to try 
out.

At 01:59 PM 9/26/2005, you wrote:
>I haven't tried this method of antigen retrieval, but TEG buffer is
>Tris-EGTA buffer. The usual formula (I have seen a couple of variations) is:
>
>Distilled water 1,000 ml
>Trisma base  1.21 gram
>EGTA  0.19 gram
>
>The usual target pH is between 8.95 and 9.0
>
>EGTA is ethylene glycol bis(B-aminoethyl)-N,N,N',N' tetraacetic acid.
>
>(The capital "B" in the above name represents "beta".  My email program
>doesn't support the symbol.)
>
> > ----------
> > From:         histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Kim
> > Merriam
> > Sent:         Monday, September 26, 2005 11:34 AM
> > To:   Histonet
> > Subject:      [Histonet] Low temp Antigen Retrieval for Bone Sections
> >
> > Hello All,
> >
> > I will be performing some IHC on mouse femurs (about 12 or so different
> > antibodies).  In the past, I have had a lot of trouble keeping these
> > sections on the slides during HIER.  I did a quick search on the archives
> > and there were lots of suggestions, but nothing definitive.
> >
> > I was reading an article about "low temp AR", overnight at 60C in TEG
> > buffer, pH 9.0. Has anyone tried this method?  Also - what does TEG buffer
> > stand for?
> >
> > Any help would be greatly appreciated.
> >
> > Kim
> >
> >
> > Kim Merriam
> > Novartis
> > Cambridge, MA
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)