Antibodies to Mouse Macrophages - FocusOn 045

F4/80 Antigen

Clones BM8 and CI:A3-1 recognise the F4/80 antigen, a 120-160 kDa glycoprotein containing seven EGF-like domains at the N-terminus, an RGD (Arg-Gly-Asp) integrin binding motif and significant homology to the transmembrane 7 (Tm7) hormone receptor family at the C-terminus. The molecular structure suggests multiple ligands. Possibly, F4/80 is involved in macrophage adhesion within certain tissues, combined with receptor signaling following peptide-ligand interaction. The antigen is expressed by most macrophages and macrophage precursors beyond M-CFC and increases upon maturation. F4/80 is widely used for the detection of tissue macrophages. Activated macrophages and eosinophils express low levels. Schaller et al. (2002) showed that BM8 antibody is the only macrophage marker that is able to distinguish non-destructive from destructive inflammation processes in the pancreas and has been shown to be a unique histological marker of the progression from peri-insulitis to ß-cell destruction and diabetes in a mouse diabetes model.

Fig. 1: Mouse spleen frozen sections stained with BM8 antibody to F4/80 Cat.-No. BM4007

Fig. 2: Mouse liver sections stained with BM8 antibody to F4/80 Cat.-No. BM4007

Fig. 3: Mouse spleen sections stained with Cl:A3-1 antibody to F4/80 Cat.-No. BM4008A

MOMA Clones

The clone MOMA-1 and MOMA-2 are widely used mouse macrophage subpopulation markers.

MOMA-1 (CD169) is a useful marker for the identification of macrophage subpopulations in various organs, mostly characterised by a high level of non-specific esterase expression. The staining is particularly noteworthy with the metallophilic macrophages adjacent to the marginal zone of the spleen. The marker is also very suitable for diffe-rentiation of non-metallophilic marginal zone macrophages as detected by ER-TR9. In addition, MOMA-1 detects macrophages at inflammatory sites and is po-sitive with Kupffer cells.

MOMA-2 is a useful marker for the broad detection of monocytes and macrophages in all mouse strains. In combination with the anti F4/80 markers BM8 or Cl:A3-1 it allows a precise characterisation of tissue fixed macrophages in various organs. The antibody stains a mature macrophage subset, monocytes and a few precursors in bone marrow. Dendritic cells show low to intermediate expression. MOMA-2 is predominantly expressed in the cytoplasm, but is also present on the cell surface. The major drawback of both clones is that they cannot be used on paraffin sections like BM8 or Cl:A3-1.

Fig. 4: Staining of mouse spleen using MOMA-1 Cat.-No. SM066P. Note the typical staining of metallophilic macrophages in the marginal zone

Antibody to Macrophage Precursor Cells (Ly-6C) Clone ER-MP20

ER-MP20 antibody is useful for the detection of macrophage precursor cells in the mid-stage development stage (Ly-6C, late CFU-M, monoblasts and monocytes) (s. Fig.5.). It is highly suitable for the detection of monocytes in bone marrow samples by FACS. ER-MP20 also identifies activated macrophages in inflammatory tissues where the simultaneous use of the murine pan-macrophage marker BM8 (anti F4/80) is recommended. ER-MP20 antibody also detects a wide range of endothelial cells.

Fig. 5: Staining of mouse liver using antibody clone ER-MP20 Cat.-No. BM4019

Mouse Macrosialin (CD68)

Macrosialin is a heavily glycosylated transmembrane protein of 87-115 kDa, which is specifically expressed by tissue macrophages, Langerhans cells and at low levels by dendritic cells. Macrosialin is the murine homologue of the human macrophage glycoprotein CD68. Both are members of the lysosomal-associated membrane protein (lamp) family which are located predominantly within the cells and can be detected by flow cytometry using cell permeabilisation. CD68 antibody clone FA-11 detects surface macrosialin at low levels in resident mouse peritoneal macrophages which can be enhanced with thioglycolate stimulation. It is the optimal tool for detection of cells of the macrophage/monocyte lineage by flow cytometry and is available with different labels.

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Key References

1

Schaller, E. et al.: Inactivation of the F4/80 glycoprotein in the mouse germ line.
Mol Cell Biol, 2002; 22: 8035-43.

3

Kraal, G. et al.: Macrophages in T and B cell compartments and other tissue macrophages recognized by monoclonal antibody MOMA-2.
Scand J Immunol, 1987; 26: 653-661.

Further readings

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