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      RB705 Mouse Anti-Bcl-6
      RB705 Mouse Anti-Bcl-6
      Flow cytometric analysis of Bcl-6 expression in Human Ramos cells.  Cells from the Human Ramos (Burkitt's lymphoma, ATCC® CRL-1596™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) followed by washing and intracellular staining with BD Horizon™ RB705 Mouse IgG1, κ Isotype Control (Cat. No. 570261; dashed line histogram) or BD Horizon™ RB705 Mouse Anti-Bcl-6 antibody (Cat. No. 570617/570705; solid line histogram). The histogram showing Bcl-6 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      RB705 Mouse Anti-Bcl-6
      Multicolor flow cytometric analysis of Bcl-6 expression in Mouse B lymphocytes.  BALB/c Mouse mesenteric lymph node cells were stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 553092), BD Horizon™ BUV395 Rat Anti-Mouse CD4 (Cat. No. 568375), BD Horizon™ BV421 Hamster Anti-Mouse CD95 (Cat. No. 562633), and BD Pharmingen™ APC-H7 Rat Anti-Mouse IgD (Cat. No. 565348) antibodies. 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™ RB705 Mouse Anti-Bcl-6 antibody (Cat. No. 570617/570705). The bivariate pseudocolor density plot showing the correlated expression of IgD versus CD95/Fas on B cells was derived from CD4-B220+ gated events with the forward and side light-scatter characteristics of intact lymphocytes (Left Plot). Germinal center (GC) B cells were identified as CD95/Fas-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 Plot) derived from gated cell subsets show intracellular Bcl-6 staining levels for Mouse 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 Cell Analyzer System and FlowJo™ Software.
      RB705 Mouse Anti-Bcl-6 RB705 Mouse Anti-Bcl-6
      Flow cytometric analysis of Bcl-6 expression in Human Ramos cells.  Cells from the Human Ramos (Burkitt's lymphoma, ATCC® CRL-1596™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) followed by washing and intracellular staining with BD Horizon™ RB705 Mouse IgG1, κ Isotype Control (Cat. No. 570261; dashed line histogram) or BD Horizon™ RB705 Mouse Anti-Bcl-6 antibody (Cat. No. 570617/570705; solid line histogram). The histogram showing Bcl-6 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Multicolor flow cytometric analysis of Bcl-6 expression in Mouse B lymphocytes.  BALB/c Mouse mesenteric lymph node cells were stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 553092), BD Horizon™ BUV395 Rat Anti-Mouse CD4 (Cat. No. 568375), BD Horizon™ BV421 Hamster Anti-Mouse CD95 (Cat. No. 562633), and BD Pharmingen™ APC-H7 Rat Anti-Mouse IgD (Cat. No. 565348) antibodies. 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™ RB705 Mouse Anti-Bcl-6 antibody (Cat. No. 570617/570705). The bivariate pseudocolor density plot showing the correlated expression of IgD versus CD95/Fas on B cells was derived from CD4-B220+ gated events with the forward and side light-scatter characteristics of intact lymphocytes (Left Plot). Germinal center (GC) B cells were identified as CD95/Fas-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 Plot) derived from gated cell subsets show intracellular Bcl-6 staining levels for Mouse 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 Cell Analyzer System 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/test
      604, 12053
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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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 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.

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      Product Notices

      1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      6. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      7. 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).
      8. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      9. 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.
      10. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      11. 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.
      12. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
      13. For U.S. patents that may apply, see bd.com/patents.
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      570617 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.

      570617 Rev. 1
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      570617 Rev.1
      Citations & References
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      View product citations for antibody "570617" on CiteAb

      Development References (10)

      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. Choi YS, Kageyama R, Eto D, et al. ICOS receptor instructs T follicular helper cell versus effector cell differentiation via induction of the transcriptional repressor Bcl6.. Immunity. 2011; 34(6):932-46. (Clone-specific: Flow cytometry). View Reference
      3. 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
      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. Fazilleau N, McHeyzer-Williams LJ, Rosen H, McHeyzer-Williams MG. The function of follicular helper T cells is regulated by the strength of T cell antigen receptor binding. Nat Rev Immunol. 2009; 10(4):375-384. (Biology). View Reference
      8. 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
      9. Klein U, Dalla-Favera R. Germinal centres: role in B-cell physiology and malignancy. Nat Rev Immunol. 2008; 8(1):22-33. (Biology). View Reference
      10. 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 (10) View Less
      570617 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.