ED Clone Antibodies for Studying Rat Macrophage Subtypes

ED Clone Antibodies for Studying Rat Macrophage Subtypes

Bio-Rad is the primary manufacturer of the complete range of ED clone antibodies, offering unrivaled quality at the most competitive prices.

ED1 (CD68): Pan-macrophage marker

Our monoclonal antibody MCA341 is the widely used pan-macrophage marker recognizing CD68, a single chain glycoprotein of 90-110 kDa. CD68 staining is predominantly found on the lysosomal membrane of myeloid cells, where there is also weak cell surface expression. The majority of tissue macrophages express CD68, while peripheral granulocytes only stain weakly. Expression levels of CD68 correlate with the phagocytic activity of macrophages.

MCA341 is available in several formats, including purified, biotinylated, and a wide range of fluorescently-labeled conjugates. The CD68, ED1 clone antibody has been validated for use in flow cytometry, immunoprecipitation, Western blotting, and immunohistology on frozen and paraffin sections.

ED2 (CD163)

MCA342 recognizes CD163, a 175 kDa cell surface glycoprotein present on resident rat macrophages. This antibody is a useful tool for staining synovial lining and Kupffer cells in the liver. It can also be used to discriminate between thymic cortical macrophages (ED2 +ve) and thymic medullary macrophages (ED2 -ve). However, CD163 is not expressed by monocytes, alveolar macrophages, or microglial cells.

MCA342 is available in purified, biotinylated, and several fluorescently-labeled formats. The CD163, ED2 clone antibody has been successfully used for flow cytometry, immunoprecipitation, Western blotting, and immunohistology on frozen and paraffin sections.

ED3 (CD169)

MCA343 recognizes the rat CD169 cell surface antigen, a 185 kDa molecule expressed by macrophages predominately confined to lymphoid organs. CD169 is a receptor for glycoconjugates containing sialic acid. The ED3 antibody stains marginal zone macrophages and marginal metallophils in the spleen very strongly.

CD169 is also expressed in macrophages in auto-immune diseased tissues, such as those associated with multiple sclerosis. As a result, it is being investigated for the targeted delivery of drugs, toxins, and antigens via sialoadhesion-specific immunoconjugates as a therapeutic strategy to control cells expressing CD169. No other cell types have been shown to be positive for ED3 (CD169) staining or expression, including healthy tissue.

MCA343 has been validated for flow cytometry, immunoprecipitation, and immunohistology on frozen sections.

ED5: Follicular Dendritic Cell Marker

MCA530R staining reveals a web-like pattern in B cell follicles which represents the follicular dendritic cells. Mesangial cells in the kidney are also positive for staining by the ED5 clone antibody. The ED5 antibody (MCA530R) has been certified for immunohistology on frozen sections.

ED7 and ED8 (CD11b): Macrophages, Monocytes, Dendritic Cells, and Granulocytes

The ED7 clone antibody (MCA618R) recognizes a membrane antigen, CD11b, on rat macrophages, monocytes, dendritic cells, and granulocytes. The recognized antigen is a 160 kDa/95 kDa heterodimer belonging to the CD11b/CD18 family of adhesion molecules and is also known as Mac-1 or CR3. ED8 (MCA619) is believed to recognize a second epitope from the same molecule.

ED7and ED8 clone antibodies have been shown detect small ramified microglia in the central nervous system as well as cilia of the bronchus epithelium. No other cell types are positive for staining. Both ED7 and ED8 can be used to induce homotypic aggregation of granulocytes.

ED7 (MCA618) is certified for flow cytometry, functional studies, and immunohistology on frozen sections. ED8 (MCA619) is available in purified, FITC, and RPE-labeled formats and has also been validated for flow cytometry and frozen sections.

ED9 (CD172a): Signal Regulatory Protein

MCA620 recognizes rat CD172a, also known as signal regulatory protein (SIRP). CD172a is expressed by myeloid cells and neurons. The ED9 clone antibody binds to a different epitope on SIRP than the CD172a OX-41 (MCA274) antibody and has been reported to block the interaction of CD172a with CD47.

MCA620 is available in purified, FITC, and RPE-labeled versions and has been qualified for use in flow cytometry, immunoprecipitation, Western blotting, and immunohistology on frozen sections.

ED10: Reticular Elements in Secondary Lymphoid Tissue

MCA732 detects reticular elements in the T cell areas of the spleen, lymph nodes, and Peyer’s patches, as well as those in bronchus and gut associated lymphoid tissues (BALT and GALT). In the thymus, a reticular staining pattern is observed in the capsule and medulla. In non-lymphoid organs, such as liver, lung, and small intestine, some ED10 positive reticular are found. No staining is observed in skin, kidney, brain, pancreas or heart.

MCA732 is available as ascites and has been qualified to work on frozen sections.

ED11: Complement C3

Biochemical studies have revealed that the ED11 clone antibody, MCA733R, recognizes complement factor C3. Like anti-ED10, it targets reticular elements in the T cell areas and B cell follicles of spleen, lymph nodes, and Peyer’s patches, as well as bronchus and gut associated lymphoid tissues. In the thymus a reticular pattern is observed throughout the cortex capsule, while the medulla is negative for ED10 signal. In vivo studies have shown that ED11 blocks immune complex trapping on follicular dendritic cells.

In non-lymphoid organs, positive staining is only seen in reticular elements of the small intestine; no staining is observed in liver, lung, skin, kidney, brain, pancreas, or heart.

MCA733R is certified for use in ELISA and for immunohistology on frozen sections.

Key References*

ED1, ED2, & ED3

  1. Dijkstra, C.D. et al. (1985) The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3.
    Immunology 54: 589-599

This reference has been cited in more than 100 publications, click here to see the complete list.

ED5 & ED6

  1. Jeurissen, S.H.M and Dijkstra, C.D. (1986) Characteristics and functional aspects of non lymphoid cells in rat germinal centres, recognized by two monoclonal antibodies ED5 and ED6.
    Eur. J. Immunol. 16: 562-568.

ED7, ED8, & ED9

  1. Damoiseaux, J.G. et al. (1989) Heterogeneity of macrophages in the rat evidenced by variability in determinants: two new anti-rat macrophage antibodies against a heterodimer of 160 and 95 kd (CD11/CD18).
    J. Leukoc. Biol. 46: 556-564
  2. Damoiseaux J.G. et al. (1989) Rat bone marrow and monocyte cultures: influence of culture time and lymphokines on the expression of macrophage differentiation antigens.
    J Leukoc Biol. 46:246-53.

ED10 & ED11

  1. Van den Berg, T.K. et al. (1989) The heterogeneity of the reticulum of rat peripheral lymphoid organs identified by monoclonal antibodies.
    Eur. J. Immunol. 19: 1747-1756.
  2. Van den Berg T.K. et al. (1992) Selective inhibition of immune complex trapping by follicular dendritic cells with monoclonal antibodies against rat C3.
    Eur. J. Immunol. 22: 957-962.

*More references are available on the individual ED clone antibody datasheets.