Mouse anti-Human CD163 Monoclonal Antibody | anti-CD163 antibody

CD163

Cat. Num. AAA370129

• Reactivity: Human
• Synonyms: CD163; Monoclonal Antibody; anti-CD163 antibody

• Host: Mouse
• Reactivity: Human
• Clonality: Monoclonal
• Isotype: IgG1
• Clone Number: 10D6
• Form/Format: CD163 is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

• Control: Placenta, Tonsil, Lymph Node, Inflamed Tissue, H. Pylori
• Localization: Cytoplasmic, Membranous
• Immunogen: Recombinant protein encoding of domains 1-4 of the N-terminal region of human CD163.
• Reactivity Note: Paraffin, Frozen
• Precautions: 1. This product contains <0.1% sodium azide (NaN3) as a preservative. Ensure proper handling procedures are used with this reagent.; 2. Always wear personal protective equipment such as laboratory coat, goggles and gloves when handling reagents.; 3. Dispose of unused solution with copious amount of water.; 4. Do not ingest reagent. If reagent is ingested, seek medical advice immediately.; 5. Avoid contact with eyes. If contact occurs, flush with large quantities of water.; 6. Follow safety precautions of the heating device used for epitope retrieval (TintoRetriever Pressure Cooker or similar).; 7. For additional safety information refer to Safety Data Sheet for this product.; 8. For complete recommendations for handling biological specimens, please refer to the CDC document, “Guidelines for Safe Work Practices in Human and AMDL”.
• Specimen Preparation: Paraffin sections: The antibody can be used on formalin-fixed paraffin-embedded (FFPE) tissue sections. Ensure tissue undergoes appropriate fixation for best results. Pre-treatment of tissues with heat-induced epitope retrieval (HIER) is recommended using ImmunoDNA Retriever with Citrate, ImmunoDNA Retriever with EDTA or ImmunoDNA Digestor. See reverse side for complete protocol. Tissue should remain hydrated via use of Immuno/DNA Washer solutions.; Frozen sections and cell preparations: The antibody can be used for labeling acetone-fixed frozen sections and acetone-fixed cell preparations.
• Staining Procedure: 1. Cut and mount 3-5 micron formalin-fixed paraffin-embedded tissues on positively charged slides such as Hydrophilic Plus Slides.; 2. Air dry for 2 hours at 58° C.; 3. Deparaffinize, dehydrate and rehydrate tissues.; 4. Subject tissues to heat induced epitope retrieval (HIER) using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate or EDTA.; 5. Any of three heating methods may be used:; a. TintoRetriever Pressure Cooker or Equivalent; Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA, and place on trivet in the pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high. Incubate for 15 minutes. Open and immediately transfer slides to room temperature.; b. TintoRetriever PT Module or Water Bath Method; Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA at 95°-99° C. Incubate for 30-60 minutes.c. Conventional Steamer Method; Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a steamer, cover and steam for 30-60 minutes.; 6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.; 7. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.; 8. Wash slides with ImmunoDNA washer or DI water.; 9. Continue IHC staining protocol. Wash slides between each step with ImmunoDNA washer solution.
• Mounting Protocols: For detailed instructions using biodegradable permanent mounting media such as XyGreen PermaMounter or organic solvent based resin such as PermaMounter.
• Product Limitations: Due to inherent variability present in immunohistochemical procedures (including fixation time of tissues, dilution factor of antibody, retrieval method utilized and incubation time), optimal performance should be established through the use of positive and negative controls. Results should be interpreted by a qualified medical professional.
• Preparation and Storage: Store at 2 to 8 degree C in the dark. (Control Slides: Store at 20-25 degree C. )

Dilution Information

Immunohistochemistry (IHC)

Abbreviated Immunohistochemical Protocol

Related Product Information for anti-CD163 antibody

CD163 is a protein that in humans is encoded by the CD163 gene. CD163 is the high affinity scavenger receptor for the hemoglobin-haptoglobin complex and in the absence of haptoglobin - with lower affinity - for hemoglobin alone. CD163 is expressed exclusively on the cell surface of human monocytes and macrophages that evolve predominantly in the late phase of inflammation, and is, therefore, very useful for macrophage-phenotyping. A soluble form of the receptor exists in plasma, commonly named sCD163, which is upregulated in a large range of inflammatory diseases including liver cirrhosis, type 2 diabetes, atherosclerosis, macrophage activation syndrome, Gaucher’s disease, sepsis, HIV infection, rheumatoid arthritis and Hodgkin Lymphoma.

CD163 positivity by IHC can be seen in histiocytes, gut, Kupffer cells, a few alveolar macrophages, the main population of macrophages in the placenta, and in varying degrees in macrophages in inflamed tissue including tumor tissue, depending on the inflammatory stage. Red-pulp, not white-pulp, macrophages in the spleen and cortical macrophages of the thymus are also positive for this marker. CD163 has been found to be helpful in distinguishing synovial macrophages from synovial intimal fibroblasts in the setting of rheumatoid arthritis, with superior specificity for macrophages than CD68, which does not discriminate between these cell types. It also has been confirmed in previous reports of having a prognostic role of tumor-infiltrating macrophages in classical Hodgkin’s Lymphoma. Increased levels of CD163 have been detected in patients with microbial infections and myelomonocytic leukemias and studies have confirmed the fact that CD163 expression is limited to leukemias with monocytic differentiation. Another recent study showed that all 5 cases of synovial-type giant cell tumors of the spinal column were positive for CD163.

NCBI and Uniprot Product Information

• NCBI GI #: 5107945
• UniProt Accession #: Q86VB7; Q07898; Q07899; Q07900; Q07901; Q2VLH7; C9JIG2
• NCBI Official Full Name: CD163
• UniProt Protein Name: Scavenger receptor cysteine-rich type 1 protein M130
• Protein Family: Scavenger receptor cysteine-rich type 1 protein
• UniProt Gene Name: CD163
• UniProt Synonym Gene Names: M130
• UniProt Entry Name: C163A_HUMAN

NCBI Description

The protein encoded by this gene is a member of the scavenger receptor cysteine-rich (SRCR) superfamily, and is exclusively expressed in monocytes and macrophages. It functions as an acute phase-regulated receptor involved in the clearance and endocytosis of hemoglobin/haptoglobin complexes by macrophages, and may thereby protect tissues from free hemoglobin-mediated oxidative damage. This protein may also function as an innate immune sensor for bacteria and inducer of local inflammation. Alternatively spliced transcript variants encoding different isoforms have been described for this gene. [provided by RefSeq, Aug 2011]

Uniprot Description

Function: Acute phase-regulated receptor involved in clearance and endocytosis of hemoglobin/haptoglobin complexes by macrophages and may thereby protect tissues from free hemoglobin-mediated oxidative damage. May play a role in the uptake and recycling of iron, via endocytosis of hemoglobin/haptoglobin and subsequent breakdown of heme. Binds hemoglobin/haptoglobin complexes in a calcium-dependent and pH-dependent manner. Exhibits a higher affinity for complexes of hemoglobin and multimeric haptoglobin of HP*1F phenotype than for complexes of hemoglobin and dimeric haptoglobin of HP*1S phenotype. Induces a cascade of intracellular signals that involves tyrosine kinase-dependent calcium mobilization, inositol triphosphate production and secretion of IL6 and CSF1. Isoform 3 exhibits the higher capacity for ligand endocytosis and the more pronounced surface expression when expressed in cells. Ref.7 Ref.9 Ref.10 Ref.12 Ref.18After shedding, the soluble form (sCD163) may play an anti-inflammatory role, and may be a valuable diagnostic parameter for monitoring macrophage activation in inflammatory conditions. Ref.7 Ref.9 Ref.10 Ref.12 Ref.18

Subunit structure: Interacts with CSNK2B. Ref.9

Subcellular location: Soluble CD163: Secreted Ref.7 Ref.8. Cell membrane; Single-pass type I membrane protein. Note: Isoform 1 and isoform 2 show a lower surface expression when expressed in cells. Ref.7 Ref.8

Tissue specificity: Expressed in monocytes and mature macrophages such as Kupffer cells in the liver, red pulp macrophages in the spleen, cortical macrophages in the thymus, resident bone marrow macrophages and meningeal macrophages of the central nervous system. Expressed also in blood. Isoform 1 is the lowest abundant in the blood. Isoform 2 is the lowest abundant in the liver and the spleen. Isoform 3 is the predominant isoform detected in the blood. Ref.7 Ref.10 Ref.18

Induction: Induced by anti-inflammatory mediators such as glucocorticoids, interleukin-6/IL6 and interleukin-10/IL10; suppressed by proinflammatory mediators like bacterial lipopolysaccharides (LPS), IFNG/IFN-gamma and TNF. Ref.7 Ref.8

Domain: The SRCR domain 3 mediates calcium-sensitive interaction with hemoglobin/haptoglobin complexes. Ref.15

Post-translational modification: A soluble form (sCD163) is produced by proteolytic shedding which can be induced by lipopolysaccharide, phorbol ester and Fc region of immunoglobulin gamma. This cleavage is dependent on protein kinase C and tyrosine kinases and can be blocked by protease inhibitors. The shedding is inhibited by the tissue inhibitor of metalloproteinase TIMP3, and thus probably induced by membrane-bound metalloproteinases ADAMs.Phosphorylated

Probable. Ref.9

Miscellaneous: Intravenous lipopolysaccharide (LPS) produces a rapid rise of sCD163 in plasma of patient as it induces metalloproteinase-mediated shedding from monocytes surface. Long-term LPS infusion finally increases expression of the membrane-bound form on circulating monocytes.The soluble form (sCD163) in plasma is a novel parameter in diseases affecting macrophage function and monocyte/macrophage load in the body. The concentration of sCD163 is probably reflecting the number of macrophages of the 'alternative macrophage activation' phenotype with a high CD163 expression playing a major role in dampening the inflammatory response and scavenging components of damaged cells. This has initiated a number of clinical studies for evaluation of sCD163 as a disease marker in inflammatory conditions e.g. infection, autoimmune disease, transplantation, atherosclerosis and cancer.

Sequence similarities: Contains 9 SRCR domains.

Caution: It is uncertain whether Met-1 or Met-6 is the initiator.

Biophysicochemical propertiesKinetic parameters:KM=2.0 nM for hemoglobin/haptoglobin of HP*1S phenotype Ref.10KM=0.2 nM for hemoglobin/haptoglobin of HP*1F phenotype

Sequence caution: The sequence CAA80541.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.The sequence CAA80542.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.The sequence CAA80543.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.The sequence CAA80544.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.The sequence CAB45233.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.