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      BUV496 Mouse Anti-Bcl-6
      BUV496 Mouse Anti-Bcl-6
         Bcl-6 expression in Ramos cells (Left Panel). Human Ramos (Burkitt's lymphoma, ATCC CRL-1596) cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) followed by intracellular staining with either BD Horizon™ BUV496 Mouse IgG1, κ Isotype Control (Cat. No. 612949; dashed line histogram) or BD Horizon™ BUV496 Mouse Anti-Human Bcl-6 antibody (Cat. No. 567752/567753, solid line histogram). The fluorescence histogram showing the expression of Bcl-6 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells.    Bcl-6 expression in mouse B cells (Middle and Right Panels). Mouse mesenteric lymph node cells were stained with PE Rat Anti-Mouse B220 (Cat. No. 553090), Alexa Fluor™ 700 Rat Anti-Mouse CD4 (Cat. No. 557956), BD Horizon™ BV786 Hamster Anti-Mouse Fas/CD95 (Cat. No. 740906), and BD Horizon™ BUV395 Rat Anti-Mouse IgD (Cat. No. 565988). Cells were washed, resuspended in RPMI 1640 medium, and fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049). Cells were permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885), followed by intracellular staining with BD Horizon™ BUV496 Mouse Anti-Bcl-6 antibody. The pseudocolor density plot showing the correlated expression of IgD versus Fas/CD95 on B cells was derived from CD4-B220+ gated events with the light-scatter characteristics of intact lymphocytes (Middle Panel). Germinal center (GC) B cells were identified as Fas/CD95-positive B cells that expressed low levels of IgD (IgDloCD95/Fas+) whereas non-GC B cells primarily expressed intermediate to high levels of IgD and little or no CD95/Fas. Histograms (Right Panel) derived from gated cell subsets show intracellular Bcl-6 staining levels for GC B cells (solid line) and non-GC B cells (dashed line).    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 and FlowJo™ software.
      BUV496 Mouse Anti-Bcl-6
         Bcl-6 expression in Ramos cells (Left Panel). Human Ramos (Burkitt's lymphoma, ATCC CRL-1596) cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) followed by intracellular staining with either BD Horizon™ BUV496 Mouse IgG1, κ Isotype Control (Cat. No. 612949; dashed line histogram) or BD Horizon™ BUV496 Mouse Anti-Human Bcl-6 antibody (Cat. No. 567752/567753, solid line histogram). The fluorescence histogram showing the expression of Bcl-6 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells.    Bcl-6 expression in mouse B cells (Middle and Right Panels). Mouse mesenteric lymph node cells were stained with PE Rat Anti-Mouse B220 (Cat. No. 553090), Alexa Fluor™ 700 Rat Anti-Mouse CD4 (Cat. No. 557956), BD Horizon™ BV786 Hamster Anti-Mouse Fas/CD95 (Cat. No. 740906), and BD Horizon™ BUV395 Rat Anti-Mouse IgD (Cat. No. 565988). Cells were washed, resuspended in RPMI 1640 medium, and fixed with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049). Cells were permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885), followed by intracellular staining with BD Horizon™ BUV496 Mouse Anti-Bcl-6 antibody. The pseudocolor density plot showing the correlated expression of IgD versus Fas/CD95 on B cells was derived from CD4-B220+ gated events with the light-scatter characteristics of intact lymphocytes (Middle Panel). Germinal center (GC) B cells were identified as Fas/CD95-positive B cells that expressed low levels of IgD (IgDloCD95/Fas+) whereas non-GC B cells primarily expressed intermediate to high levels of IgD and little or no CD95/Fas. Histograms (Right Panel) derived from gated cell subsets show intracellular Bcl-6 staining levels for GC B cells (solid line) and non-GC B cells (dashed line).    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 and FlowJo™ software.
      Product Details
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      BD Horizon™
      BCL6; B-cell lymphoma 6 protein; LAZ3; Laz-3, ZBTB27, ZNF51
      Human (QC Testing), Mouse (Tested in Development)
      Mouse IgG1, κ
      Human Bcl-6 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      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.
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      Recommended Assay Procedures

         BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead 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).

         Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

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      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. 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).
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. BD Horizon Brilliant Ultraviolet 496 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
      6. 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.
      7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
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      567752 Rev. 1
      Antibody Details
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      K112-91

      The K112-91 monoclonal antibody specifically binds to Bcl-6. Bcl-6 was first identified as a proto-oncogene frequently deregulated by chromosomal translocations in non-Hodgkin B-cell lymphomas. It is a nuclear transcriptional repressor of the BTB/POZ zinc-finger family of transcription factors. In addition to its roles in cancer, Bcl-6 plays important roles in the differentiation of normal cells including B cells, thymocytes, CD4+ or CD8+ T cells. Bcl-6 is highly expressed in germinal center B cells, where it promotes the germinal center reaction by inducing proliferation and inhibiting the DNA-damage response. Bcl-6 has been identified as a key factor in promoting the differentiation of CD4+ follicular T helper (Tfh) cells that are involved in promoting germinal center formation and providing help to B cells. The interplay of Bcl-6 and another transcriptional repressor, Blimp-1, is thought to be critical in defining the results of both B-cell and T-cell differentiation.

      567752 Rev. 1
      Format Details
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      BUV496
      The BD Horizon Brilliant™ Ultraviolet 496 (BUV496) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 496-nm. BUV496, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 500-nm (e.g., 515/30-nm bandpass filter). The acceptor dye can be excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BUV496
      Ultraviolet 355 nm
      350 nm
      496 nm
      567752 Rev.1
      Citations & References
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      View product citations for antibody "567752" on CiteAb

      Development References (8)

      1. Baumjohann D, Okada T, Ansel KM. Cutting Edge: Distinct Waves of BCL6 Expression during T Follicular Helper Cell Development. J Immunol. 2011; 187(5):2089-2092. (Clone-specific: Flow cytometry). View Reference
      2. Chung Y, Tanaka S, Chu F, et al. Follicular regulatory T cells expressing Foxp3 and Bcl-6 suppress germinal center reactions. Nat Med. 2011; 17(8):983-988. (Clone-specific: Flow cytometry). View Reference
      3. Crotty S, Choi YS, Kageyama R, et al. ICOS receptor instructs T follicular helper cell versus effector cell differentiation via induction of the transcriptional repressor Bcl6. Immunity. 2011; 34:1-15. (Clone-specific: Flow cytometry). View Reference
      4. Crotty S, Johnston RJ, Schoenberger SP. Effectors and memories: Bcl-6 and Blimp-1 in T and B lymphocyte differentiation. Nat Immunol. 2010; 11(2):114-120. (Biology). View Reference
      5. Crotty S. Follicular Helper CD4 T Cells (Tfh). Annu Rev Immunol. 2011; 29(1):621-663. (Biology). View Reference
      6. Eto, D., C. Lao, et al. IL-21 and IL-6 are critical for different aspects of B cell immunity and redundantly induce optimal follicular helper CD4 T cell (Tfh) differentiation. PLoS ONE. 2011; 6(3):e17739. (Clone-specific: Flow cytometry). View Reference
      7. Johnston RJ, Poholek AC, DiToro D, et al. Bcl6 and Blimp-1 are reciprocal and antagonistic regulators of T follicular helper cell differentiation.. Science. 2009; 325(5943):1006-10. (Biology). View Reference
      8. Nurieva RI, Chung Y, Martinez GJ, et al. Bcl6 mediates the development of T follicular helper cells. Science. 2009; 325(5943):1001-1005. (Biology). View Reference
      View All (8) View Less
      567752 Rev. 1

       

      Please refer to Support Documents for Quality Certificates


      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.