I'm confused. The F4/80 antibody I know (clone BM8) is a rat monoclonal that stains macrophages and macrophage derived lineages in mice beautifully. Kuppfer cells, in spleen, skin, lymph nodes using an anti-rat secondary that has been pre-absorbed against mouse. For peroxidase or immunoflouresence and people all over research use F4/80 on mouse tissues. I think the problem is with what and how are your microglia "activated". Not ALL macrophage markers work on microglia in different states because they are not the exact flavor of a card carrying macrophage. Try a liver or LN and see if you get no staining.
Ray Koelling
PhenoPath Labs
Seattle, WA
-------------- Original message --------------
From: rschoon@email.unc.edu
> F4/80 is a RAT specific marker for activated macrophages so I am not to
> surprised that nothing is staining in the mouse tissues. I've used
> this antibody with great success on rat in the past. Suggest that you
> search the web maybe stating here:
>
> Activated microglia in cortex of mouse models of mucopolysaccharidoses
> I and IIIB Kazuhiro Ohmi*,, David S. Greenberg*,, Kavitha S. Rajavel*,
> Sergey Ryazantsev*, Hong Hua Li*, and Elizabeth F. Neufeld*,,§,¶ *
> Department of Biological Chemistry and Brain Research Institute, David
> Geffen School of Medicine, and § Molecular Biology Institute,
> University of California, Los Angeles, CA 90095
>
> Contributed by Elizabeth F. Neufeld, December 20, 2002
>
> -N-Acetylglucosaminidase deficiency (mucopolysaccharidosis IIIB, MPS
> IIIB) and -L-iduronidase deficiency (MPS I) are heritable lysosomal
> storage diseases; neurodegeneration is prominent in MPS IIIB and in
> severe cases of MPS I. We have obtained morphologic and molecular
> evidence for the involvement of microglia in brain pathology of mouse
> models of the two diseases. In the cortex, a subset of microglia
> (sometimes perineuronal) consists of cells that are probably
> phagocytic; they have large storage vacuoles, react with MOMA-2
> (monoclonal antibody against macrophages) and Griffonia simplicifolia
> isolectin IB4, and stain intensely for the lysosomal proteins Lamp-1,
> Lamp-2, and cathepsin D as well as for GM3 ganglioside. MOMA-2-positive
> cells appear at 1 and 6 months in MPS IIIB and MPS I mice,
> respectively, but though their number increases with age, they remain
> sparse. However, a profusion of cells carrying the macrophage
> CD68/macrosialin antigen appear in the cortex of both mouse models at 1
> month. mRNA encoding CD68/macrosialin also increases at that time, as
> shown by microarray and Northern blot analyses. Ten other transcripts
> elevated in both mouse models are associated with macrophage functions,
> including complement C4, the three subunits of complement C1q, lysozyme
> M, cathepsins S and Z, cytochrome b558 small subunit,
> macrophage-specific protein 1, and DAP12. An increase in IFN- and IFN-
> receptor was observed by immunohistochemistry. These functional
> increases may represent activation of resident microglia, an influx and
> activation of blood monocytes, or both. They show an inflammatory
> component of brain disease in the two MPS, as is known for many
> neurodegenerative disorders.
>
> Robert Schoonhoven
> Scientist, Pathology
> MPI Research
> --------------------------------------------------------------------------------
> K.O. and D.S.G. contributed equally to this work.
>
> "Dear colleagues,
> is anybody there who has experiences with a F4/80 antibody? It is
> supposed to label "activated" microglia. Unfortunatly I don't get a
> specific staining in mice.
> Does anyone have an idea?
> Thanks a lot,
>
> F. Neff"
>
>
>
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