CD163 Monoclonal Antibody (eBioGHI/61 (GHI/61)), Super Bright™ 600, eBioscience™, Invitrogen™

Beschreibung

Description: The eBioGHI/61 monoclonal antibody reacts with human CD163 (Haemoglobin Scavenger Receptor, M130), a member of the macrophage scavenger receptor family. CD163 is expressed by tissue macrophages, circulating blood monocytes, and some reports suggest that it may also be expressed by haematopoietic progenitor cells. CD163 functions as a high affinity scavenger receptor for the complex of haemoglobin and haptoglobin which forms to clear haemoglobin from the blood. The high affinity for CD163 is specific for the haemoglobin-haptoglobin complex, whereas haemoglobin or haptoglobin display no to low affinity toward CD163. Upon ligand binding, CD163 initiates signal transduction which leads to the production of IL-6 and IL-10. Furthermore, it has been demonstrated that cytokines such as IL-6 and IL-10 can induce expression of CD163, whereas proinflammatory stimuli, such as LPS or TNF alpha, can cause shedding of a soluble form of CD163, whose function is unclear. Applications Reported: This eBioGHI/61 antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBioGHI/61 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 μL (1.0 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product No. SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product No. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. CD163 (M130 antigen, Ber-Mac3, Ki-M8, SM4) is a 130 kDa membrane glycoprotein, a member of the scavenger receptor cysteine-rich superfamily, and a receptor for the hemoglobin-haptoglobin complex. CD163 protects tissues from free hemoglobin-mediated oxidative damage, and may play a role in the uptake and recycling of iron, via endocytosis of hemoglobin/haptoglobin and subsequent breakdown of heme. CD163 is expressed exclusively on the cell surface of human monocytes and macrophages that evolve predomitly in the late phase of inflammation. Specifically, CD163 is present on all circulating monocytes and most tissue macrophages except those found in the mantle zone and germinal centers of lymphoid follicles, interdigitating reticulum cells and Langerhan′s cells. CD163 is present on all CD14 positive monocytes, most CD64 positive monocytes, and shows higher expression on CD16 positive monocytes. CD163 is upregulated on mononuclear phagocytes by IL-10, IL-6 and dexamethasone. Lipopolysaccharide (LPS) and phorbol myristate acetate (PMA) both induce shedding of CD163 from the cell surface into plasma or cell supernatant. CD163 binds hemoglobin/haptoglobin complexes in a calcium-dependent and pH-dependent manner, and exhibits a higher affinity for complexes of hemoglobin and multimeric haptoglobin of HP1F phenotype than for complexes of hemoglobin and dimeric haptoglobin of HP1S phenotype. Further, CD163 also induces a cascade of intracellular signals that involves tyrosine kinase-dependent calcium mobilization, inositol triphosphate production and secretion of IL6 and CSF1.