[Histonet] To detect injected mouse antibody inside mouse tissue

[koellingr <@t> comcast.net]

Amy, 

In my opinion the answer is, Yes, certainly possible.  Also in my opinion the answer is No, probably impossible.  Even with a Yahoo address I'm assuming that this is a research/biotech model and project.  In a previous life, when quietly gagged, I went about this on numerable occassions.  Sometimes extraordinarily successfully and sometimes complete failures.  Done at both light IHC level and with EM IHC.  Depends on your specific antibody molecule, how many target receptors on a given cell, pharmacokinetics of the injected antibody, making sure it gets to target and in sufficient quantity, not sequestered inappropriately, how long animal sacrificed post-injection.  Probably 5 other variables, each with nuances so just impossible to answer generally in this venue for your specific problem.  There are several papers on the subect if you look them up but they are specific procedures for a very specific model.  If successful, results are spectacular.  More often than not, you spend months on something that in retrospect was probably not doable in the first place after you sit and really think it through.  I once got bitten to the tune of several months work and at the end we found out something about the glycosylation of the antibody that would never allow us to localize it appropriately so we had been searching for something we could never find by IHC. 



Ray 



Raymond Koelling 

PhenoPath Labs 

Seattle, WA 
----- Original Message ----- 
From: "Amy Lee" <amylee779 <@t> yahoo.com> 
To: "histonet" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Thursday, January 21, 2010 2:19:35 PM GMT -08:00 US/Canada Pacific 
Subject: [Histonet] To detect injected mouse antibody inside mouse tissue 

Hello, 
  
I have a question regarding detecting injected mouse antibody in mouse tissue by IHC. 
Now I have mouse liver paraffin section. This mouse was injected mouse antibody. I was asked to locate this antibody. This is a mouse chimeric antibody. Variable regions from human antibody was combined with mouse IgG2a heavy chain and kappa light chain constant regions. I am thinking using rabbit anti-human IgG, F(ab) fragment specific. Do you think it's doable or you have any good suggestion? When this mouse antibody bind to antigen inside liver through this human variable region, my F(ab) fragment specific antibody can still bind to it? 
  
Hope I describ my questions clearly. 
  
Thank you in advance for any help! 
  
Amy 



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