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PE Mouse Anti-Human BAFF (BLyS/CD257)
PE Mouse Anti-Human BAFF (BLyS/CD257)

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 PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-BAFF (BLyS/CD257) antibody (Cat. No. 567738; solid line histogram) at 1 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.

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 PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dashed line histogram) or PE Mouse Anti-BAFF (BLyS/CD257) antibody (Cat. No. 567738; solid line histogram) at 1 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.

Product Details
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BD Pharmingen™
B-cell-activating factor; B-lymphocyte stimulator; BAFF; BLYS; CD257; DTL; TALL1; THANK; TNFSF13B
Human (QC Testing)
Mouse IgG1, κ
Human BAFF Recombinant Protein
Flow cytometry (Routinely Tested)
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. An isotype control should be used at the same concentration as the antibody of interest.
567738 Rev. 1
Antibody Details
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H32-411

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.

567738 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567738 Rev.1
Citations & References
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Development References (9)

  1. 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
  2. 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
  3. Mackay F, Schneider P. Cracking the BAFF code.. Nat Rev Immunol. 2009; 9(7):491-502. (Biology). View Reference
  4. 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
  5. 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). View Reference
  6. 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
  7. 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
  8. 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
  9. 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
View All (9) View Less
567738 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.