[Histonet] Endogenous Peroxide Treatment - Immunohistochemistry

[Jacqui Detmar]

Hi Marilyn.  I work on mouse placenta - also very bloody - and I find
that I get the best peroxidase quenching when I put the slides in 3%
H202 in methanol for 20-30 minutes.  Also, for most of my antibodies, I
can do the quenching step after the biotinylated secondary antibody, but
BEFORE adding the ABC solution.  Just make sure the slides are well
washed after quenching.

Hope this helps,

Jacqui

Jacqui Detmar, Post-doctoral Fellow
Samuel Lunenfeld Research Institute, room 876
Mount Sinai Hospital
600 University Avenue
Toronto, ON, Canada
M5G 1X5

phone:   416-586-4800 x2451/x2290
fax:        416-586-8588
email:     detmar <@t> mshri.on.ca


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Marilyn
Johnson
Sent: Saturday, March 15, 2008 2:27 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Endogenous Peroxide Treatment - Immunohistochemistry

Hi Histonetters,
For routine tissues, I use 3% hydrogen peroxide for 10 mins. to remove
RBC's.
Presently, I am trying to stain liver tissue, which stores more of the
blood components.
There seems to be background staining that masks blood cells and
vessels.
I've tried using 5% hydrogen peroxide solution for 10 mins. and there is
no improvement.
Are there other solutions that can be used instead of hydrogen peroxide?
Any replies would be greatly appreciated.
Thank you in advance.

Marilyn Johnson
Alberta Agriculture
Food and Safety Division
Edmonton, Alberta, Canada.
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