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 amylee...@yahoo.com
To: histonet histonet@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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