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 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.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Pacific Blue™ is a trademark of Life Technologies Corporation.
- 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.
Companion Products
The MAC2-158 monoclonal antibody (also known as Clone MAC 2-158) specifically binds to human CD163. CD163 is also known as Scavenger receptor cysteine-rich type 1 protein M130 (M130), SCARI1 (SR-I1), or Hemoglobin scavenger receptor (HbSR). CD163 is an ~130 kDa type I transmembrane glycoprotein comprised of an extracellular domain with nine cysteine-rich (SRCR) scavenger receptor class B domains followed by a transmembrane region and a short cytoplasmic tail. CD163 is expressed on most peripheral blood monocytes, tissue macrophages, and a subset of dendritic cells. CD163 serves as a high affinity receptor for hemoglobin and haptoglobin and mediates endocytosis of hemoglobin and haptoglobin complexes by macrophages. This scavenging function may protect tissues from hemoglobin-mediated oxidative damage and contribute to the uptake and recycling of iron. CD163 can also reportedly bind to (TNF-a)-like weak inducer of the apoptosis (TWEAK) protein and some pathogenic bacteria. A cleaved, soluble form of CD163 can reportedly play an anti-inflammatory role and serve as a marker for macrophage activation in inflammatory responses. High-affinity binding of the MAC2-158 antibody to CD163 is reportedly unaffected by extracellular calcium levels. This clone can be used to measure CD163 expression in freshly drawn whole blood samples stabilized with commonly used anticoagulants, eg, EDTA, citrate or heparin.
Development References (8)
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Bover LC, Cardó-Vila M, Kuniyasu A, et al. A previously unrecognized protein-protein interaction between TWEAK and CD163: potential biological implications.. J Immunol. 2007; 178(12):8183-94. (Biology). View Reference
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Etzerodt A, Moestrup SK. CD163 and inflammation: biological, diagnostic, and therapeutic aspects. Antioxid Redox Signal. 2013; 18(17):2352-2363. (Biology). View Reference
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Fabriek BO, van Bruggen R, Deng DM, et al. The macrophage scavenger receptor CD163 functions as an innate immune sensor for bacteria.. Blood. 2009; 113(4):887-92. (Biology). View Reference
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Högger P, Sorg C. Soluble CD163 inhibits phorbol ester-induced lymphocyte proliferation.. Biochem Biophys Res Commun. 2001; 288(4):841-3. (Biology). View Reference
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Jensen AL, Collins J, Shipman EP, Wira CR, Guyre PM, Pioli PA. A subset of human uterine endometrial macrophages is alternatively activated. Am J Reprod Immunol. 2012; 68(5):374-386. (Clone-specific: Flow cytometry). 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). View Reference
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Morganelli PM, Guyre PM. IFN-gamma plus glucocorticoids stimulate the expression of a newly identified human mononuclear phagocyte-specific antigen.. J Immunol. 1988; 140(7):2296-304. (Immunogen: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
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PrabhuDas MR, Baldwin CL, Bollyky PL, et al. A Consensus Definitive Classification of Scavenger Receptors and Their Roles in Health and Disease.. J Immunol. 2017; 198(10):3775-3789. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.