[Histonet] Endogenous Peroxide Treatment -IHC

From:"Margaryan, Naira"

Hi histonetters,

I fully agree with Isaac.

We all know that the liver always gives a background even if 4 steps of
blocking were used (H2O2, Avidin, Biotin, Protein Block).

Why in TMAs always we found a liver as a control sample, which gives us
false result???????????????


Message: 2
Date: Sat, 15 Mar 2008 11:29:08 -0700
From: "Isaac Milos" 
Subject: RE: [Histonet] Endogenous Peroxide Treatment -
To: "Marilyn Johnson" ,
Content-Type: text/plain;	charset="us-ascii"

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!


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