Re: Miracles Needed

From:Gayle Callis <>

Neil Hand wrote a publication in Journal of Histotechnology using stringent
antigen retrieval for tissue embedded in methyl methacrylate on microtomed
sections. It requires total removal of MMA with warm xylene, and then
pressure cooker retrieval.  He can be reached at Neil Hand
<> to help you out.  I believe the
article was written in 1998.  He is considered an expert in this area and
has a panel of 100 antibodies or so that he has done with MMA with
wonderful staining.  

MMA must be removed or you will not be able to do IHC, it is hydrophobic,
and antibodies, etc cannot get to antigenic sites.  

At 10:18 AM 10/4/00 -0400, you wrote:
>Dear Histonetters,
>I have inherited a project of 2% paraformaldehyde/2%
>glutaraldehyde/ruthenium red fixed rat tibia embedded at low temperature in
>MMA.  I hope someone out there is immunostaining bone and can give me
>Basically, I need two miracles:  
>1)  BrDU   Can anyone tell me why I can not retrieve this antigen in MMA? 
>(I have tried steam, boiling citrate
>buffer, autoclave, pepsin digestion)   A couple specimens were embedded in
>epon/araldite and can be successfully stained with a Becton-Dickinson
>primary antibody.   
>2)  TGF-beta1   Can anyone suggest a primary antibody for TGF-beta1 to be
>used in this same rat tibia tissue?
>As you can see I really need your collective experience.  Thanks in
>Judy Strauss
>Institute for Human Performance
>SUNY Upstate Medical University
>Department of Orthopedic Surgery
>Syracuse, New York
Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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