Re: IH on Liver Tissue
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From: | Gayle Callis <uvsgc@msu.oscs.montana.edu> |
To: | histonet@pathology.swmed.edu |
Reply-To: | |
Content-Type: | text/plain; charset=us-ascii |
There is a peroxidase quench that will block ALL forms of peroxidase,
totally. This is the glucose oxidase method.
Hsu H-M, et al Am J Pathology 188:209-217,1984
Andrew SM and Jasani G Histchem J 19:426-430, 1987
Vector provided me with the exact protocol
>Date: Mon, 10 Jul 2000 14:43:03 -0400
>From: Jeff Crews <jcrews@organo.com>
>Subject: Re: IH on Liver Tissue
>To: kkdulany@unmc.edu, histonet@pathology.swmed.edu
>
> With extremely bloody tissue like spleen or liver, it is sometimes
> impossible to quench all of the peroxidase present in the blood. Try
> an alkaline-phoshatase detection system.
>
> Jeffrey Crews, HTL (ASCP)
> Organogenesis, Inc.
>
>
>______________________________ Reply Separator
_________________________________
>Subject: IH on Liver Tissue
>Author: <kkdulany@unmc.edu> at internet
>Date: 07/10/2000 11:03 AM
>
>
>How can you successfully perform immunohistochemistry on slides of liver
>tissue using horse-radish peroxidase and DAB? How do you quench the
>endogenous peroxidase in the tissue so everything doesn't turn brown? I've
>tried 3, 10 and 15 % H2O2 in both methanol and di water for 20 min. up to 1
>hour with no success. The tissues are formalin fixed and embeded in
>paraffin. We then used a pH 6.0 citrate buffer/microwave antigen retrieval
>system for our primary Ab detection. Even with all that the endogenous
>enzyme caused the false positive with the DAB. Anyone have any
>suggestions?
>Thank you for all replies.
>Karen in Omaha
>
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Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303
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