RE: IH on Liver Tissue
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From: | "Nocito, Joseph" <joseph_nocito@srhc.iwhs.org> |
To: | "'kkdulany@unmc.edu'" <kkdulany@unmc.edu>, histonet@pathology.swmed.edu |
Reply-To: | |
Content-Type: | text/plain |
Karen
liver & kidney have a high amount of endogenous biotin in them. Several
immuno companies have an avidin-biotin blocking kit to use as a
pretreatment. You place avidin on the slides for 15 minutes, rinse in PBS,
then place biotin on for 15 minutes, rinse, then continue with your usual
immuno procedure. I usually perform this treatment after blocking with
Methanol/Peroxide and before applying the primary. Good luck.
Joe Nocito, B.S., HT(ASCP)QIHC
Histology Supervisor
Christus Santa Rosa Hospitals
San Antonio, Texas
> -----Original Message-----
> From: kkdulany@unmc.edu [SMTP:kkdulany@unmc.edu]
> Sent: Monday, July 10, 2000 11:04 AM
> To: histonet@pathology.swmed.edu
> Subject: IH on Liver Tissue
>
> 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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