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Flow cytometric analysis of CD163 expression on human peripheral blood monocytes. Human whole blood was stained with either BD Horizon™ BV786 Mouse IgG1, κ Isotype Control (Cat. No. 563330; dashed line histogram) or BD Horizon™ BV786 Mouse Anti-Human CD163 antibody (Cat. No. 568221; solid line histogram) at 1 µg/test. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD163 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV786 Mouse Anti-Human CD163
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
The GHI/61 monoclonal antibody specifically binds to human CD163. CD163 is also known as Scavenger receptor cysteine-rich type 1 protein M130 (M130), Hemoglobin scavenger receptor and Macrophage-associated antigen. CD163 is a 110-130 kDa transmembrane glycoprotein. CD163 is a monocyte/macrophage-restricted antigen expressed on the majority of tissue macrophages and peripheral blood monocytes. CD163 belongs to the scavenger receptor superfamily. Its expression on monocytes is upregulated upon cellular activation. CD163 expression reportedly changes on monocytes and macrophages as these cells differentiate. This finding suggests a role for this molecule in the differentiation and/or regulation of monocyte and macrophage function. CD163 may play a role in the clearance and endocytosis of hemoglobin and haptoglobin complexes by macrophages.
It has been reported (Maniecki et al., 2011) that the presence of calcium impacts the binding affinity of clone GHI/61 to CD163. There is a variation in detecting CD163 positive monocytes when the cells are prepared with different anticoagulants, where heparin was observed to have the highest inhibitory effect on clone GHI/61.
Development References (4)
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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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Law SK, Micklem KJ, Shaw JM. A new macrophage differentiation antigen which is a member of the scavenger receptor superfamily. Eur J Immunol. 1993; 23(9):2320-2325. (Biology). View Reference
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Maniecki MB, Etzerodt A, Moestrup S, Møller J, Graversen J. Comparative assessment of the recognition of domain-specific CD163 monoclonal antibodies in human monocytes explains wide discrepancy in reported levels of cellular surface CD163 expression. Immunobiology. 2011; 216(8):882-890. (Biology). View Reference
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Pulford K, Micklem K, McCarthy S, Cordell J, Jones M, Mason DY. A monocyte/macrophage antigen recognized by the four antibodies GHI/61, Ber-MAC3, Ki-M8 and SM4. Immunology. 1992; 75(4):588-595. (Immunogen: Blocking, Flow cytometry, Immunoaffinity chromatography, Immunohistochemistry, Immunoprecipitation). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.