Macrophages antibody

Catalog No. ABIN111879

Product Details

Target: Macrophages

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: Un-conjugated

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

Specificity: Monoclonal antibody 25F9 is associated with fully differentiated tissue macrophages both in normal and diseased tissues, particularly also in the late stage of an inflammation. Antigen Distribution Isolated Cells: Absent on freshly isolated monocytes and other blood cells, present on 40-50 % of human monocytes after 6-7 days in culture, also positive on some melanoma and carcinoma cell lines. Tissue Sections: Kupffer cells, histiocytes (skin), macrophages of the thymus, in the germinal centres of lymph nodes and spleen, in mamma carcinoma, melanoma, osteocarcinoma and gastric cancer, eczema, sarcoidosis, BCG granuloma, synovial lining cells, tuberculoid leprosy: no expression in lepromatous leprosy. Antibody BM4022 reacts with Human Mature Macrophages and Monocytes. Other Species (tested so far): Subpopulation of macrophages in Rhesus monkey, reactive with pig alveolar macrophages and Kupffer cells.

Cross-Reactivity (Details): Species reactivity (tested):Human.

Purification: Affinity Chromatography.

Immunogen: Cultured human monocytes. Remarks: The antigen is a 86 kDa membrane protein, the epitope has not been further characterized.

Clone: 25F9

Isotype: IgG1

Application Details

Application Notes: Immunohistochemistry on Frozen Sections: 1 μg/mL (1/200). Immunohistochemistry on Paraffin Sections: 2 μg/mL (1/100). Proteinase K pretreatmentfor antigen retrieval recommended. Suggested Positive Control: Human tonsil. The antibody is suitable for staining macrophages from bronchial lavage fluids and similartechniques. It is very useful for macrophage phenotyping, particularly for the classificationof late inflammatory stages (together with the anti calprotectin clone 27E10 and the antiCD163 clone 5C6-FAT). It is used in tissue sections and in smears, for the characterization of tumorous tissues andthe monitoring of macrophage cell cultures.
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 mouse IgG+IgM(H+L) minimal-cross reaction to human) for 1h in a humid chamber8. Wash in PBS9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS11. Counterstain. Protocol with formalin-fixed, paraffin-embedded sections: The whole procedure is performed at room temperature1. Deparaffinize and rehydrate tissue section2. Incubate the tissue section with proteinase K for 7min. 3. Wash in distilled water4. Block endogenous peroxidase5. Wash in PBS6. Block with 10% normal goat serum in PBS for 30min. in a humid chamber

Restrictions: For Research Use only

Handling

Reconstitution: Restore with 0.5 mL distilled water.

Concentration: 0.2 mg/mL

Buffer: 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: This product is photosensitive and should be protected from light

Storage: 4 °C

Storage Comment: Store the antibody undiluted Prior to and following reconstitution at 2-8 °C. DO NOT FREEZE!

Target Details

Target: Macrophages

Background: Synonyms: Macrophage marker