This Monoclonl antibody ER-MP20 is useful for the detection of macrophage precursor cells in the mid-stage development stage (late CFU-M, monoblasts and monocytes). It is ideally 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 antibody BM4007) is recommended. ER-MP20 also detects a wide range of endothelial cells. Antigen Distribution on Isolated cells: In bone marrow cells the antigen is found on monoblasts and late CFU-M cells as well as on monocytes. It is also found on granulocytes and a subpopulation of lymphocytes in the peripheral blood. Granulocytic cells show a dull, and monocytic cells a bright antigen surface expression. Lymphoid cells express the antigen only very weakly. Thus, in the bone marrow three useful FACS windows can be defined for cell sorting purposes. Antigen Distribution on Tissue Sections: The antigen is found on macrophage precursor subpopulations in the bone marrow and hemopoietic islands of the lymphoid organs, and in the spleen. Endothelial cells of small vessels in various organs also stain positive with ER-MP20. Activated macrophages in inflammatory tissues also express the ER-MP20-related antigen.
Cross-Reactivity (Details)
Species reactivity (tested):Mouse (Macrophage precursor cells, endothelial cells).
Purification
Affinity Chromatography
Immunogen
Mouse macrophage cell lines. Remarks: Antigen/Epitope: The antigen is a glutaraldehyde (0.05%) and paraformaldehyde (1%) resistant 14kD surface protein which is very similar to Ly-6C and may be analogous to Human CD59. It is inducible by IFN-alpha, IFN-beta and IFN-gamma.
Ly-6C
Reactivity: Mouse
FACS
Host: Rat
Monoclonal
Monts 1
PE
Application Notes
Immunohistochemistry on Frozen Sections: 0.2 μg/mL (1/1000)Immunohistochemistry on Paraffin Sections: 0.4 μg/mL (1/500). Proteinase K pretreatmentfor antigen retrieval is recommended. Suggested Positive Control: Mouse spleen. Has been described to work in FACS. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Restrictions
For Research Use only
Reconstitution
Restore with 0.5 mL distilled water.
Concentration
0.2 mg/mL
Buffer
Stock solution containing PBS, pH 7.2 with 5 mg/mL BSA as a stabilizer and 0.09 % Sodium Azide as a preservative
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Avoid repeated freezing and thawing. This product is photosensitive and should be protected from light
Storage
4 °C/-20 °C
Storage Comment
Store the antibody at 2-8 °C for 2 months or (in aliquots) at -20 °C for longer.
Ly-6C is a member of the Ly-6 multigene family of type V glycophosphatidylinositol-anchored cell surface proteins. It is expressed on bone marrow cells, monocytes/macrophages, neutrophils, endothelial cells, and T-cell subsets. Mice with the Ly-6.2 allotype (e.g., AKR, C57BL, C57BR, C57L, DBA/2, PL, SJL, SWR, 129) have subsets of CD4+Ly-6C+ and CD8+Ly-6C+ cells, while Ly-6.1 strains (e.g., A, BALB/c, CBA, C3H/He, DBA/1, NZB) have only CD8+Ly-6C+ lymphocytes. Ly-6C may play a role in the development and maturation of lymphocytes.Synonyms: Ly-6C2, Ly6C2, Lymphocyte antigen 6C2, Mouse Macrophage Marker