Re: [Histonet] blocking endogenous peroxidase when using ABC stainingand DAB

From:" Katri Tuomala"

Hi Rebekah,
It could be you are dealing with something else than endogenous peroxidase.
We need to know more detail of your method to try and think it through:
1.What animal is your primary antibody raised in and is it known to work on
2.What are you using to block non-specific staining?
3.What animal is your secondary antibody raised in?
4. Are you working with formalin fixed, paraffin embedded or frozen
There are a lot of techs working on animal tissues on HistoNet. If you
provide more information, I'm sure help will be forthcoming...


Katri Tuomala
Department of Pathology
St.Joseph's Health Care
----- Original Message ----- 
Sent: Wednesday, June 09, 2004 10:18 AM
Subject: [Histonet] blocking endogenous peroxidase when using ABC
stainingand DAB

> Hi,I'm working as a lab technician at UF making using
> immunohistochemistry (ABC method and then DAB, specifically) to
> find out where eNOS is located in sheep placenta tissue. However,
> I seem to be both not getting staining on the endothelial cells
> (which is weird since I'm using eNOS antibody) and getting a lot
> of staining due to endogenous peroxidase activity. I've already
> tried putting the slides in a solution of 1:10 30%hydrogen
> peroxide to methanol for as long 40 min, and it still doesn't seem
> to quench the endogenous peroxidase. Anyone have any suggestions
> as to other methods of quenching it?
> --
> "You are a child of the universe, no less than the trees and the
> stars
> You have a right to be here and whether or not it is clear to you,
> no doubt the universe is unfolding as it should. Therefore be at
> peace with G-d, whatever you conceive Him to be. And whatever your
> labors and aspirations,in the noisy confusion of life, keep peace
> in your soul.-Max Ehrmann,"Desiderata"
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