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Multiparameter flow cytometric analysis using BD OptiBuild™ RB613 Mouse Anti-Human CD163 antibody (Cat. No. 759278) on Human peripheral blood. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
BD OptiBuild™ RB613 Mouse Anti-Human CD163
Regulatory Statusの凡例
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
推奨アッセイ手順
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- CF™ is a trademark of Biotium, Inc.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
関連製品
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 (5)
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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. (Clone-specific: Flow cytometry, Western blot). 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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Pulford K, Micklem K, Thomas J, Jones M, Mason DY. A 72-kD B cell-associated surface glycoprotein expressed at high levels in hairy cell leukaemia and plasma cell neoplasms. Clin Exp Immunol. 1991; 85(3):429-435. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.