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Flow cytometric analysis of BAFF (BLyS/CD257) expression on human U937 cells. Cells from the human U937 (Histiocytic Lymphoma, ATCC CRL-1593.2) cell line were stained with APC Mouse IgG1, κ Isotype Control (Cat. No. 554681; dashed line histogram) or APC Mouse Anti- Human BAFF (BLyS/CD257) antibody (Cat. No. 568052/568053; solid line histogram) at 2 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing BAFF (BLyS/CD257) expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ APC Mouse Anti-Human BAFF (BLyS/CD257)
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 cell 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.
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Companion Products
The H32-411 monoclonal antibody specifically recognizes B-cell activating factor (BAFF) which is also known as B-lymphocyte stimulator protein (BLyS), TNF and ApoL-related leukocyte expressed ligand 1 (TALL1), Dendritic cell-derived TNF-like molecule (DTL) and CD257. BAFF is a type II transmembrane glycoprotein that is encoded by TNFSF13B (TNF superfamily member 13b). BAFF can be expressed on B cells, macrophages, and dendritic cells as well as on tumor cells. Membrane and soluble forms of BAFF have been described. In its soluble form, BAFF can serve as a potent B cell growth factor in costimulation assays, suggesting its biological role in monocyte-driven B cell activation. BAFF has also been linked to B cell hyperplasia and autoimmunity.
Development References (9)
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Hase H, Kanno Y, Kojima M, et al. BAFF/BLyS can potentiate B-cell selection with the B-cell coreceptor complex.. Blood. 2004; 103(6):2257-65. (Biology). View Reference
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Litinskiy MB, Nardelli B, Hilbert DM, et al. DCs induce CD40-independent immunoglobulin class switching through BLyS and APRIL.. Nat Immunol. 2002; 3(9):822-9. (Biology). View Reference
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Mackay F, Schneider P. Cracking the BAFF code.. Nat Rev Immunol. 2009; 9(7):491-502. (Biology). View Reference
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Moore PA, Belvedere O, Orr A, et al. BLyS: member of the tumor necrosis factor family and B lymphocyte stimulator.. Science. 1999; 285(5425):260-3. (Biology). View Reference
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Mukhopadhyay A, Ni J, Zhai Y, Yu GL, Aggarwal BB. Identification and characterization of a novel cytokine, THANK, a TNF homologue that activates apoptosis, nuclear factor-kappaB, and c-Jun NH2-terminal kinase.. J Biol Chem. 1999; 274(23):15978-81. (Biology: Flow cytometry). View Reference
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Nardelli B, Belvedere O, Roschke V, et al. Synthesis and release of B-lymphocyte stimulator from myeloid cells.. Blood. 2001; 97(1):198-204. (Biology). View Reference
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Scapini P, Nardelli B, Nadali G, et al. G-CSF-stimulated neutrophils are a prominent source of functional BLyS.. J Exp Med. 2003; 197(3):297-302. (Biology). View Reference
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Schneider P, MacKay F, Steiner V, et al. BAFF, a novel ligand of the tumor necrosis factor family, stimulates B cell growth.. J Exp Med. 1999; 189(11):1747-56. (Biology). View Reference
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Shu HB, Hu WH, Johnson H. TALL-1 is a novel member of the TNF family that is down-regulated by mitogens.. J Leukoc Biol. 1999; 65(5):680-3. (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.