Whole Mount Immunohistochemistry

jim crouch James_crouch at smtpgw.musc.edu
Mon Sep 25 13:16:41 EST 1995


In article <43kktn$oq2 at amgen.amgen.com>, DJ Kane <dkane at amgen.com> wrote:

> Does anyone have a good, working protocol for whole mount 
> immunohistochemistry (on whole mouse embryos), using anti-neurofilament 
> antibody, for example?
> 
> Thanks in advance....

First of all the most important thing is the location of the antigen. 
Inside the cell(cytoplasmic) requires the use of Triton X pre-treatment. 
Don't use triton in any of your other solutions as it may dissociate your
Ag-Ab complex.  I have had good sucess using whole mount immuno on whole
cochleas from gerbils.  Definitly get very thin sections of your embryo. 
No reason why paraffin sections won't work.  My mentors have over 400 pubs
in immunohistochemistry and we have had great success with whole mounts. 
Good luck.

Jim Crouch



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