I am looking for help with Ab staining of embryos. I have generated an
antibody that works great on westerns, however in embryos it gives extremely
weak signals (and almost no background). I suspect that the antigen needs
to be denatured in embryos (just like it is in the western blot).
I have been using a standard fixation protocol: 4% paraformaldehyde in
My question :
Is there a way to modify the fixation to include a detergent or something
which will denature my antigen ? Should I use this detergent throughout
the Ab staining protocol ? Has anyone encontered this problem before (and
solved it) ?
Any help is greatly appreciated