Macrophages (Haematopoiesis associated) Mouse Monoclonal Antibody [Clone ID: 25F9]

Specifications

Product Data
Clone Name	25F9
Applications	IHC
Recommend Dilution	Immunohistochemistry on Frozen Sections: 0.25 μg/ml (1/800). Immunohistochemistry on Paraffin Sections: 0.5 μg/ml (1/400). Proteinase K pretreatment for antigen retrieval recommended. Suggested Positive Control: Human tonsil. The antibody is suitable for staining macrophages from bronchial lavage fluids and similar techniques. It is very useful for macrophage phenotyping, particularly for the classification of late inflammatory stages (together with the anti calprotectin clone 27E10 and the anti CD163 clone 5C6-FAT). It is used in tissue sections and in smears, for the characterization of tumorous tissues and the monitoring of macrophage cell cultures.
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Immunogen	Cultured Human monocytes.
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.
Formulation	PBS, pH 7.2 State: Purified State: Lyophilized purified Ig fraction Stabilizer: 5 mg/ml BSA Preservative: 0.05% Kathon CG
Reconstitution Method	Restore with 0.5 ml distilled water.
Concentration	0.2 mg/ml
Purification	Affinity Chromatography
Conjugation	Unconjugated
Storage Condition	Store lyophilized at 2-8°C for 6 months or at -20°C long term. After reconstitution store the antibody undiluted at 2-8°C for one month  or (in aliquots) at -20°C long term. Avoid repeated freezing and thawing.
Synonyms	Macrophage marker
Note	Protocol: Protocol with frozen, ice-cold acetone-fixed sections: The whole procedure is performed at room temperature 1. Wash in PBS 2. Block endogenous peroxidase 3. Wash in PBS 4. Block with 10% normal goat serum in PBS for 30min. in a humid chamber 5. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber 6. Wash in PBS 7. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM (H+L) minimal-cross reaction to human) for 1h in a humid chamber 8. Wash in PBS 9. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 10. Wash in PBS 11. Counterstain. Protocol with formalin-fixed, paraffin-embedded sections: The whole procedure is performed at room temperature 1. Deparaffinize and rehydrate tissue section 2. Incubate the tissue section with proteinase K for 7min. 3. Wash in distilled water 4. Block endogenous peroxidase 5. Wash in PBS 6. Block with 10% normal goat serum in PBS for 30min. in a humid chamber 7. Incubate with primary antibody (dilution see datasheet) for 1h in a humid chamber 8. Wash in PBS 9. Incubate with secondary antibody (peroxidase-conjugated goat anti mouse IgG+IgM (H+L) minimal-cross reaction to human) for 1h in a humid chamber 10. Wash in PBS 11. Incubate with AEC substrate (3-amino-9-ethylcarbazol) for 12min. 12. Wash in PBS 13. Counterstain.
Reference Data

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