Thank you for your reply about the MDM2 staining.
I used citrate buffer in the Decloaker (a pressure cooker) for 5 min and
incubated with primary antibody overnight at room temperature and used a
polymer detection system (Ultravision) from LabVision for 30 min. I also
tried EDTA buffer and protease without much success.
With BondMax autostainer, I used pretreatment for 20 or 30 mininutes with
two different buffers (ER1-citrate based and ER2-high pH buffer) and ER2
gave much sharper nuclear staining. The primary incubation time was 30
minutes. The detection kit I used was Bond Polymer Refine Detection kit.
Senior Hospital Scientist
Dept of Anatomical Pathology
Concord NSW 2139
From: Gayle Callis [mailto:email@example.com]
Sent: Friday, 1 June 2007 12:38 AM
To: Young Kwun
Subject: Re: [Histonet] Help with MDM2 Protein Immunostaining
If you tell us HOW you do your protocol, then people can help with your
background staining - which can come from many sources?
At 05:53 PM 5/30/2007, you wrote:
>Does anyone in Histoland use MDM2 Protein staining for some soft tissue
>I tried Lab Vision's polyclonal antibody (1/100 dil) and Novocastra's MDM2
>(clone 1B10, 1/150 dil) with Decloaker HIER and BondMax Autostainer.
>However, my pathologist is not happy with the staining results, mainly
>because of some non-specific stains.
>Any help would be appreciated. Thank you.
>Senior Hospital Scientist
>Dept of Anatomical Pathology
>Concord NSW 2139
>Histonet mailing list
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
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