[Histonet] Endogenous Peroxide Treatment - Immunohistochemistry

[Isaac Milos]

Hi Marilyn,

If you are running your IHC stains with an HRP detection kit with a
biotinylated secondary antibody, endogenous biotin within the tissue can
cause background staining to occur.  Often times, you'll see endogenous
biotin in a tissue that is particularly bloody, such as liver, kidney,
or brain.

You can use an avidin/biotin block before secondary antibody is applied
to account for this.  You can find this block through many different
companies - mine (Cell Marque) offers a good version.  The part number
is CMX222 and it can be ordered at 1-800-665-7284.

Happy staining!

Isaac

-----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 11:27 AM
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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