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Flow cytometric analysis of CD268 (BAFF-R) expression on human peripheral blood lymphocytes. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human CD268 (BAFF-R) antibody (Cat. No. 564819/565989; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV421 Mouse Anti-Human CD268 (BAFF Receptor)
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
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349).
Product Notices
- 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.
- 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.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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 www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 11C1 monoclonal antibody recognizes CD268 which is also known as the B cell activating factor receptor (BAFF-R). CD268 is a type III transmembrane protein of approximately 184 residues. BAFF-R is one of the receptors for B-cell activating factor (BAFF), a member of the TNF family of proteins. BAFF is a key regulator for B cell differentiation and critical in regulating survival and activation of peripheral B cell populations. Experiments in the mouse model show that interaction of BAFF with BAFF-R promotes NF-κB activity. Overexpression of BAFF results in an expanded B cell compartment and autoimmunity in mice. Mice injected with BAFF-neutralizing-Fc (BAFF-R-Fc) protein showed reduced NF-κB activation, blocking BAFF-induced B cell proliferation. Reports suggest that the BAFF-BAFF-R interaction may lead to a better understanding of autoimmune disorders.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (9)
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Karpusas M, Cachero TG, Qian F, et al. Crystal structure of extracellular human BAFF, a TNF family member that stimulates B lymphocytes. J Mol Biol. 2002; 315(5):1145-1154. (Biology). View Reference
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Kayagaki N, Yan M, Seshasayee D, et al. BAFF/BLyS receptor 3 binds the B cell survival factor BAFF ligand through a discrete surface loop and promotes processing of NF-kappaB2. Immunity. 2002; 17(4):515-524. (Biology). View Reference
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Mackay F, Browning JL. BAFF: a fundamental survival factor for B cells. Nat Rev Immunol. 2002; 2(7):465-475. (Biology). View Reference
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Ng LG, Sutherland AP, Newton R, et al. B cell-activating factor belonging to the TNF family (BAFF)-R is the principal BAFF receptor facilitating BAFF costimulation of circulating T and B cells.. J Immunol. 2004; 173(2):807-17. (Immunogen: ELISA, Flow cytometry, Functional assay, Immunohistochemistry, Inhibition). View Reference
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Ramos-Medina R, Montes-Moreno S, Maestre L, et al. Immunohistochemical analysis of HLDA9 Workshop antibodies against cell-surface molecules in reactive and neoplastic lymphoid tissues. Immunol Lett. 2011; 134(2):150-156. (Clone-specific: Immunohistochemistry). View Reference
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Schneider P, Tschopp J. BAFF and the regulation of B cell survival. Immunol Lett. 2003; 88(1):57-62. (Biology). View Reference
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Thompson JS, Bixler SA, Qian F, et al. BAFF-R, a newly identified TNF receptor that specifically interacts with BAFF. Science. 2001; 293(5537):2108-2111. (Biology). View Reference
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Vidal-Laliena M, Romero X, March S, Requena V, Petriz J, Engel P. Characterization of antibodies submitted to the B cell section of the 8th Human Leukocyte Differentiation Antigens Workshop by flow cytometry and immunohistochemistry. Cell Immunol. 2005; 236(1-2):6-16. (Clone-specific: Flow cytometry). View Reference
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Yan M, Brady JR, Chan B, et al. Identification of a novel receptor for B lymphocyte stimulator that is mutated in a mouse strain with severe B cell deficiency. Curr Biol. 2001; 11(19):1547-1552. (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.