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BUV805 Mouse Anti-Bcl-6
BUV805 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 PE Rat Anti-Mouse B220 (Cat. No. 553090), FITC  Rat Anti-Mouse CD4 (Cat. No. 553047), Alexa Fluor™ 647 Hamster Anti-Mouse Fas/CD95 (Cat. No. 563647), and BD Horizon™ BV421 Rat Anti-Mouse IgD (Cat. No. 744291) 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™ BUV805 Mouse Anti-Bcl-6 antibody. The bivariate pseudocolor density plot showing the correlated expression of IgD versus Fas/CD95 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 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 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.
BUV805 Mouse Anti-Bcl-6
Flow cytometric analysis of Bcl-6 expression in Ramos cells. Human Ramos cells (Burkitt's lymphoma, ATCC CRL-1596) 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 BD Horizon™ BUV805 mIgG1, κ Isotype Control (Cat. No. 612897; dashed line histogram) or BD Horizon™ BUV805 Mouse Anti-Bcl-6 (Cat. No. 567755/567787; 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 PE Rat Anti-Mouse B220 (Cat. No. 553090), FITC  Rat Anti-Mouse CD4 (Cat. No. 553047), Alexa Fluor™ 647 Hamster Anti-Mouse Fas/CD95 (Cat. No. 563647), and BD Horizon™ BV421 Rat Anti-Mouse IgD (Cat. No. 744291) 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™ BUV805 Mouse Anti-Bcl-6 antibody. The bivariate pseudocolor density plot showing the correlated expression of IgD versus Fas/CD95 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 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 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.
Flow cytometric analysis of Bcl-6 expression in Ramos cells. Human Ramos cells (Burkitt's lymphoma, ATCC CRL-1596) 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 BD Horizon™ BUV805 mIgG1, κ Isotype Control (Cat. No. 612897; dashed line histogram) or BD Horizon™ BUV805 Mouse Anti-Bcl-6 (Cat. No. 567755/567787; 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.
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.

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. This will 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).

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. An isotype control should be used at the same concentration as the antibody of interest.
  6. BD Horizon Brilliant Ultraviolet 805 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.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. 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.
  10. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
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.

Format Details
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BUV805
The BD Horizon Brilliant™ Ultraviolet 805 (BUV805) 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 351-nm and an acceptor dye with an emission maximum (Em Max) at 803-nm. BUV805, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 805-nm (e.g., a 820/60 or a 780/60 bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV805
Ultraviolet 355 nm
351 nm
803 nm
Citations & References
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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. 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. 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

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.