RE: [Histonet] Human anti Human

From:"Patsy Ruegg"



The only problem I see with this is similar to running an anti mouse ab on
mouse tissue.  The secondary link anti human IgG will bind to all IgG in the
human tissue, not just the primary antibody, so you would need to use serum
blocks to prevent that.  I would use 10% serum from the host of the
secondary before the primary and then again before the secondary as a block.
I would also think about using some human serum in the diluents but you have
to be careful not to block all human IgG.

Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech
12635 Montview Blvd. #215
Aurora, CO 80045
720-859-4060
fax 720-859-4110
pruegg@ihctech.net
www.ihctech.net
www.ihcrg.org    
 


-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amos Brooks
Sent: Saturday, October 25, 2008 10:19 AM
To: johnsobr@zgi.com; histonet@lists.utsouthwestern.edu
Subject: [Histonet] Human anti Human

Hi,
   I should start by saying: I have not done this, but this is the way I
would approach this problem if I were to encounter it.
   First I assume you have an human antibody raised in humans right? So, you
need an antibody that will detect the human antibody like "Anti-Human IgG
(-chain specific) antibody produced in rabbit" or "Monoclonal Anti-Human
IgG (-chain specific) antibody produced in mouse". (As seen here:
http://www.sigmaaldrich.com/life-science/cell-biology/antibodies/antibody-pr
oducts.htm?TablePage=14574992.
Then you can detect the Rabbit or Mouse secondary with your usual anti
Rabbit or Mouse detection. (I would use a polymer just to minimize biotin
background.)
   If anyone sees any holes in that line of approach, I'd love to hear other
ideas. It's an interesting problem.

Best of luck,
Amos Brooks


 Message: 22
Date: Fri, 24 Oct 2008 09:57:24 -0700
From: "BJON (Brian Johnson)" 
Subject: [Histonet] Human anti Human
To: histonet@lists.utsouthwestern.edu
Message-ID: <746E68D5CBB205409B12EC32A61EE4010A2FE8E7@ned.zgi.com>
Content-Type: text/plain; charset=us-ascii

Hello

I am wondering if anyone has found, and or used, an IHC polymer kit for
a human anti human monoclonal antibody.  Beyond the polymer kit, does
anyone have any tips for using a human anti human antibody for IHC
without biotinylating it?

Thank you.

Brian Johnson


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