Req: Immunostaining for Macrophage
wayne.conlan at nrc.ca
Fri Sep 24 14:11:51 EST 1999
Are these macrophages intralumenal or interstitial? If the former, one way to
label them is simply to inject fluorescent latex microparticles (the 1um sized
ones used for checking FACS machines) intravenously and let the macs ingest
them. This works well with fixed macs like Kupffer cells and splenic macs that
are in direct contact with the blood. Beware though, when I tried to publish
these results I was told that this technique was too old-fashioned to deliniate
macs from other mononuclear cells. Instead, I was informed I'd have to prove
that the cells in question were macs using a specific monoclonal antibody.
This is plain silly of course since macs define the specificity of the Mab, not
vice versa. In this regard, most of the so-called macrophage-specific Mabs
tend to be selective rather than specific. So by the aforementioned logic, an
epithelial cell that bound a mac-specific Mab, but didn't actively phagocytose
inert microparticles would be classified as a macrophage! Hope this helps.
Michael Rosenfeld wrote:
> Greetings, scientists,
> I am desperately searching for a reliable immunostaing protocol for the
> detection of Macrophage in frozen sections of Mouse arteries. None of the
> antibodies or protocols I have tried have worked. If anyone has a method
> that works or knows of one, please let me know. Our lab is studying
> atherosclerosis in mouse models, and obviously the macrophage is a star
> player. I'm stuck until I can reliably identify and quantitate these cells
> in atherosclerotic lesions.
> Thanks in advance, with more to follow if I get this protocol working.
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