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Flow cytometric analysis of CXCL16 expression on Human peripheral blood monocytes. Human whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD OptiBuild™ BV421 Mouse Anti-Human CXCL16 antibody (Cat. No. 750901; solid line histogram) at 0.5 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histogram showing CXCL16 expression (or Ig Isotype control staining) was derived from gated events with the forward and side-light scatter characteristics of viable 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 OptiBuild™ BV421 Mouse Anti-Human CXCL16
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 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
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
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Companion Products
The 22-19-12 monoclonal antibody specifically recognizes CXC chemokine ligand 16 (CXCL16) which is also known as C-X-C motif chemokine 16, Scavenger receptor for phosphatidylserine and oxidized low density lipoprotein (SR-PSOX), or Small-inducible cytokine B16 (SCYB16). CXCL16 is a ~30 kDa single-pass type I transmembrane glycoprotein with an extracellular CXC chemokine domain and mucin-like spacer region, followed by a transmembrane region, and a cytoplasmic domain with consensus tyrosine phosphorylation and SH2 binding sites. Transmembrane CXCL16 can serve as a scavenger receptor that binds to oxidized low density lipoprotein, phosphatidylserine, or bacteria and mediates their uptake by macrophages and dendritic cells. Cell surface CXCL16 can be shed in a soluble form after proteolytic cleavage to exert its chemotactic function. CXCL16 is differentially expressed by keratinocytes, monocytes, dendritic cells, endothelial cells, and some activated T cells. CXCL16 exerts its chemotactic function by binding to and signaling through the CXCR6 (CD186) chemokine receptor that is variably expressed by effector/memory CD8+ T cells, CD4+ T cells with Th1- or Th17-like phenotypes, γδ T cells, natural killer (NK) cells, natural killer T (NKT) cells, and monocytes. The 22-19-12 antibody reportedly binds to the chemokine domain of CXCL16 and can inhibit the chemotactic activity of soluble CXCL16.
Development References (4)
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Bachelerie F, Ben-Baruch A, Burkhardt AM, et al. International Union of Basic and Clinical Pharmacology. [corrected]. LXXXIX. Update on the extended family of chemokine receptors and introducing a new nomenclature for atypical chemokine receptors.. Pharmacol Rev. 2014; 66(1):1-79. (Biology). View Reference
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Shimaoka T, Kume N, Minami M, et al. Molecular cloning of a novel scavenger receptor for oxidized low density lipoprotein, SR-PSOX, on macrophages.. J Biol Chem. 2000; 275(52):40663-6. (Biology). View Reference
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Shimaoka T, Nakayama T, Kume N, et al. Cutting edge: SR-PSOX/CXC chemokine ligand 16 mediates bacterial phagocytosis by APCs through its chemokine domain.. J Immunol. 2003; 171(4):1647-51. (Immunogen: Flow cytometry, Functional assay, Inhibition). View Reference
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Tabata S, Kadowaki N, Kitawaki T, et al. Distribution and kinetics of SR-PSOX/CXCL16 and CXCR6 expression on human dendritic cell subsets and CD4+ T cells.. J Leukoc Biol. 2005; 77(5):777-86. (Clone-specific: Flow cytometry). View Reference
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