CD163 Mouse Monoclonal Antibody [Clone ID: 5C6-FAT]

Specifications

Product Data
Clone Name	5C6-FAT
Applications	ELISA, FC, IHC, WB
Recommend Dilution	ELISA. Immunohistochemistry on Frozen Sections: 0.5 µg/ml (1/400) Immunohistochemistry on Paraffin Sections: 1 µg/ml (1/200) (Proteinase K pretreatment for antigen retrieval is recommended). Suggested Positive Control: Human placenta. Has been described to work in FACS and Western blots.
Reactivity	Human
Host	Mouse
Clonality	Monoclonal
Immunogen	Human CD163. The epitope has not been further characterized.
Specificity	Monoclonal antibody 5C6 FAT recognizes a membrane glycoprotein on monocytes and macrophages which is expressed in intermediate and late inflammatory stages. Antigen Distribution Isolated cells: Monocytes, particularly after dexamethasone treatment or after 2-5 days in culture. Does not react with lymphocytes, granulocytes or platelets. Tissue Sections: Positive staining can be observed in the skin (histiocytes), gut, Kupffer cells, few alveolar macrophages, a major population of macrophages in the placenta, varying degrees of macrophages in inflamed tissues, including tumorous tissue depending on the inflammatory stage. Red pulp, but not white pulp macrophages of the spleen, and cortical macrophages of the thymus are detected. Macrophages in the synovialis of patients with rheumatoid arthritis. In alveolar macrophages and in Kupffer cells a double staining can be observed with monoclonal antibody 25F9 (product BM4022) which is not the case in other tissues.
Formulation	Stock Solution contains PBS, pH 7.2 with 5 mg/ml BSA as a stabilizer and 0.05% Kathon as a preservative. State: Purified State: Lyophilized purified IgG fraction
Reconstitution Method	Restore with 0.5 ml distilled water.
Concentration	0.2 mg/ml (after reconstitution)
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.
Gene Name	CD163 molecule
Database Link	Entrez Gene 9332 Human UniProt ID Q86VB7
Background	CD163 is a scavenger receptor for the haemoglobin-haptoglobin complex, and is upregulated by glucocorticoids and IL-10. The extracellular portion of the receptor is regularly shed and can be found in the circulation. An important function of CD163 seems to be in the adhesion of monocytes to activated endothelial cells. CD163-positive cells include skin histiocytes, Kupffer cells, spleen macrophages of the red pulp, and some thymus macrophages. The antigen is also found abundantly in human term placenta, and regularly in acute and chronic inflammatory lesions.
Synonyms	M130, Hemoglobin scavenger receptor, 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 with Mayer’s hemalum. 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 with Mayer’s hemalum
Reference Data

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