CD163 Rat anti-Mouse, PerCP-eFluor 710, Clone: TNKUPJ, eBioscience
Rat Monoclonal Antibody
Manufacturer: eBioscience 46163182
Description: This TNKUPJ monoclonal antibody recognizes mouse CD163. CD163 is a 130kDa surface receptor expressed by certain subsets of tissue macrophages, including splenic red pulp macrophages, Kupffer cells, intestinal lamina propria macrophages and a small fraction of peritoneal macrophages. In contrast to human blood monocytes, mouse monocytes do not express CD163. Also, unlike human CD163, mouse CD163 is not as readily induced by M2 polarizing cytokines, and it is not a good marker of M2 macrophages. No common cell lines of monocytic or macrophage origin express mouse CD163. In humans, CD163 has been shown to be proteolytically cleaved and shed from the cell surface, and it acts as a soluble anti-inflammatory factor. This TNKUPJ antibody will detect CD163 on fixed and permeabilized cells allowing for staining of the intracellular pool of this receptor. Although CD163 is relatively stable to collagenase digestion, aggressive tissue dissociation protocols might potentially decrease the amount of surface CD163. In these cases intracellular detection is recommended. Applications Reported: This TNKUPJ antibody has been reported for use in flow cytometric analysis. Applications Tested: This TNKUPJ antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.25 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. 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: 488 nm; Emission: 710 nm; Laser: Blue Laser 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 ft.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.
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
|4° C, store in dark, DO NOT FREEZE!|
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