Though a large variety of antibodies raised against various proteins expressed on macrophages can be used for e.g. MHCII, CD11b etc. The best antibodies would be CD64 and/or F4/80, these proteins are generally present on all macrophages and are generally very specific to macrophages. A word of caution, during inflammation other cell subsets can also upregulate or express these markers so use multiple markers.

CD68 or F4/80 should work in this case.

I have used a F4/80 as a pan macrophage marker before and it has stained very well.

http://www.abdserotec.com/product/mac387-anti-macrophages-monocytes-antibody-mca874g.html

Dear Venkata. I use the above marker for macrophage staining in colon biopsies. It works very well.

Best

CD68 is widely used for this purpose.

We typically use either CD11b, which stains all types of myeloid cells or CD68, which stains Macs, but also some endothelial cells.

CD68.

Try CD68 or F4/80 (although F4/80 also stains eosinophils it is much higher expressed by colonic macrophages).

Thanks everyone that was very helpful

In general CD68 is the pan-macrophage marker for human. But it is also expressed by Endothelail cells and others in tissue. Depends on the origin of the tissue and what other cell types interested in detecting together with Macrophage, one should design the staining panel with appropriate makers.

We had a lot of difficulties in using anti-F4/80 and anti-CD11b antibodies on formalin-fixed or Carnoy's-fixed mouse tissues, Does anybody else have different experience and/or protocols that do work on formalin-fixed tissues?

Sophia-

The stain depends on the company you ordered your antibody from , the blocking steps you are using

I have attached a trouble shooting guide developed after working with more than two dozen antibodies in mice, pig and human formalin fixed tissue- hope that helps you

Antibody optimization guide attached to the answer of Venkata Charepali is very nice for getting general idea how to optimize antibody performance but I was wondeirng if naybody has specific positive experience with detecting myeloid markers like CD11b and F4/80 or similar markers in humans on formalin-fixed tissues.

Also, I want to note that CD14 and CD163 are detected on tumor and inflamed endotheium in addition to other cells.

Indeed there are multiple types of tissue "macrophages", and the classical bifurcation between classical markers of macrophages and dendritic cells may not apply, especially in the gut. I'm not sure about in mice, but in human gut, for example, CD14 which is known to be expressed on monocytes/macrophages but not on DCs, and DC-SIGN, which is expressed in subsets of DCs but not macrophages, can clearly be found co-localized in large numbers of cells in the human rectal mucosa (see KB Gurney et al. May 2005. J Virol, 79: 5762-5773). In addition, see EJ Soilleux et al, Mar 2002, J Leuk Biol 71:445-457 for a comprehensive analysis of markers that co-incide with DC-SIGN expression on dendritic cell and macrophage subpopulations in vitro and in vivo. The latter is an old article but has been cited more than 250 times, so I suspect it has been useful to others. Good Luck.

Yes you can use F4/80 as a common marker to identify macrophages

hello

i want to do research in looking for foam cell in umbilical artery of baby.. what kind of stain IHC i can use? CD 68 or just Oil Red O.

Thank you

I don't think anyone has specifically stated that F4/80 only works on mouse macrophages/monocytes, not on human macrophages. The human homolog of F4/80 is EMR1 and this is primarily found on human eosinophils. So, F4/80 good for mouse, for human cells CD68 is as good as any.