I would like to identify my injected human CD14 monocytes histologically in mouse hearts, however, the antibodies we tried (even the ones specifically advertized for xenograft models) cross-reacted with the endogenous mouse macrophages, so that we can't distinguish between them and our injected human cells.
The mouse hearts are embedded in Paraffin and ideally we would like to use an avidin/Biotin/DAB staining.
Thanks for any ideas, comments, suggestions! :-)
Hello Janette,
the problem you might have is that the mouse macrophages have Fc receptors and hence bind the antibodies you used via the Fc part. If this is the case, you have to block the Fc receptors on the mouse cells. Then you might improve the work with the anti-human-antibodies for detecting the human monocytes.
Good luck!
Christoph
Hello Christoph,
thanks a lot for your answer! That is exactly what we were suspecting as well since the "unspecific binding" looked very specific and was mainly seen in the area of inflammation. Therefore, we tried to block with mouse serum (before buying any of the expensive kits that do not necessarily work) but it didn't improve the staining at all.
Does anyone have good experience maybe with a commercially available FcR block?
Thanks again!
Janette
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