Ly6C antibody

Catalog No. ABIN111872

The Rat Monoclonal anti-Ly6C antibody is suitable to detect Ly6C in samples from Mouse. It has been validated for FACS, IHC (fro) and IHC (p).

Quick Overview for Ly6C antibody (ABIN111872)

Target: Ly6C (Ly-6C) (Ly-6C)

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This Ly6C antibody is un-conjugated

Application: Flow Cytometry (FACS), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Clone: ER-MP20

Product Details anti-Ly6C Antibody

Specificity: 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).

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.

Isotype: IgG2a

Application Details

Application Notes: Immunohistochemistry on Frozen Sections: 0.5 μg/mL (1/400)Immunohistochemistry on Paraffin Sections: 1 μg/mL (1/200). 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.

Protocol: Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature1. Wash in PBS2. Block endogenous peroxidase3. Wash in PBS4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber6. Wash in PBS7. Incubate with secondary antibody (peroxidase-conjugated goat anti rat IgG (H+L)minimal-cross reaction to mouse) for 1h in a humid chamber8. Wash in PBS9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS

Restrictions: For Research Use only

Handling

Reconstitution: Restore with 0.5 mL distilled water.

Concentration: 0.2 mg/mL

Buffer: Stock solution containing PBS, pH 7.2 with 10 mg/mL BSA as a stabilizer and 0.01 % Thimerosal as a preservative

Preservative: Thimerosal (Merthiolate)

Precaution of Use: This product contains thimerosal (merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C/-20 °C

Storage Comment: Store the antibody at 2-8 °C for one month or (in aliquots) at -20 °C for longer. Avoid prolonged exposure to light. Avoid freeze/thaw cycles.

Target Details for Ly6C

Target: Ly6C (Ly-6C) (Ly-6C)

Alternative Name: Ly6c

Background: 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

Gene ID: 100041546

NCBI Accession: NP_001092687

UniProt: P09568