Re: hydrogen peroxide treatment on IHC

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From:Jamie Erickson <>,
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I think there are a few factors to consider. 1. concentration or % of H2O2 2. tissue Frozen or Paraffin also Type: spleen is very peroxidase rich. 3. Time in H2O2. WE us a Ventana TechMate 500 with H2O2 at 3% for around 5 minutes I have tried mouse and monkey tissue(s) and they seem to be fine the morphology is still good on frozens. So that's my opinion on this but the best way to find out is to try out your tissue with 3 control : positive tissue, Isotype control, and a PBS control for endogenous peroxidase.

That's my take on it, hope it helps


>>> <> 06/06 6:48 PM >>>
We currently treat some of our slides, such as kappa and B and T cell 
markers, with hydrogen peroxide prior to staining to reduce the background=20
staining that we were getting.  If anyone else does this, do you know if 
there is a time limit that you should not exceed in the hydrogen peroxide? =20
Currently we are immersing 10-15 minutes in the hydrogen peroxide before 
loading onto a Ventana automated stainer.  I wanted to know if an increase in 
time would  #1-reduce more of the background, #2- just be a waste of time, or 
#3 would damage the tissue and results.  Thanks in advance for a response.

Jennifer Macias, HT (ASCP)
Forum Health
Western Reserve Care System
Youngstown, Ohio

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