Re: [Histonet] F4/80

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 -------------- 

> 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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