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      RY610 Hamster Anti-Helios
      RY610 Hamster Anti-Helios
      Two-color flow cytometric analysis of Helios expression      Left Panel: Human peripheral blood mononuclear cells (PBMCs) were stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424) and fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with either BD Horizon™ RY610 Hamster IgG1, κ Isotype Control (Cat. No. 571251; Left Plot) or BD Horizon™ RY610 Hamster Anti-Helios antibody (Cat. No. 571235/571299; Right Plot) at 0.06 µg/test.    Right Panel: Mouse splenic leukocytes were stained with BD Pharmingen™ APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set. The cells were then stained with either Hamster IgG1, κ Isotype Control (Left Plot) or BD Horizon™ RY610 Hamster Anti-Helios antibody (Right Plot) at 0.06 µg/test. The bivariate pseudocolor density plots show the correlated expression of Helios (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact Human peripheral blood lymphocytes or splenic leukocytes.    Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      RY610 Hamster Anti-Helios
      Two-color flow cytometric analysis of Helios expression      Left Panel: Human peripheral blood mononuclear cells (PBMCs) were stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424) and fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with either BD Horizon™ RY610 Hamster IgG1, κ Isotype Control (Cat. No. 571251; Left Plot) or BD Horizon™ RY610 Hamster Anti-Helios antibody (Cat. No. 571235/571299; Right Plot) at 0.06 µg/test.    Right Panel: Mouse splenic leukocytes were stained with BD Pharmingen™ APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set. The cells were then stained with either Hamster IgG1, κ Isotype Control (Left Plot) or BD Horizon™ RY610 Hamster Anti-Helios antibody (Right Plot) at 0.06 µg/test. The bivariate pseudocolor density plots show the correlated expression of Helios (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact Human peripheral blood lymphocytes or splenic leukocytes.    Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Horizon™
      IKAROS family zinc finger protein 2; ANF1A2; HELIOS; ZNF1A2; ZNFN1A2
      Human (QC Testing), Mouse (Tested in Development)
      Armenian Hamster IgG
      Mouse Helios Peptide
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      22779, 22807
      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. 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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. 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.
      8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      9. 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.
      10. CF™ is a trademark of Biotium, Inc.
      11. For U.S. patents that may apply, see bd.com/patents.
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      571235 Rev. 1
      Antibody Details
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      22F6

      The 22F6 monoclonal antibody specifically binds to mouse and human Helios. Helios is a member of the Ikaros family of zinc-finger transcription factors, which play important roles in hematopoietic cell development and tumor suppression. Helios expression is restricted to the earliest stages of embryonic hematopoiesis, a variety of epithelial tissues and is notably increased in thymic-derived regulatory CD4+Foxp3+ T (Treg) cells. Its high expression levels in Treg cells are independent from Foxp3 and are believed to contribute, along with other transcription factors, to the phenotypic stability of natural regulatory T cells. Accordingly, it has been demonstrated that Helios directly stimulates Foxp3 transcription while it inhibits ll2 gene expression, contributing for the maintenance of cellular anergy.  Helios is also differentially expressed during negative and positive selection in the thymus, marking CD4+ autoreactive cells for deletion. Helios may possibly play roles in T cell activation, since it is upregulated in Th2 and Tfh cells. Despite these roles in T cell development and function, Helios genetic ablation in mice revealed no significant abnormalities in Treg or other T cell subsets. This finding suggests that other Ikaros family members may play redundant roles.

      571235 Rev. 1
      Format Details
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      RY610
      The BD Horizon RealYellow™ 610 (RY610) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 610-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY610 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY610 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE-CF594.
      altImg
      RY610
      Yellow-Green 561 nm
      557 nm
      610 nm
      571235 Rev.1
      Citations & References
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      View product citations for antibody "571235" on CiteAb

      Development References (10)

      1. Baine I, Basu S, Ames R, Sellers RS, Macian F. Helios induces epigenetic silencing of IL2 gene expression in regulatory T cells. J Immunol. 2013; 190(3):1008-1016. (Biology). View Reference
      2. Cai Q, Dierich A, Oulad-Abdelghani M, Chan S, Kastner P. Helios deficiency has minimal impact on T cell development and function. J Immunol. 183(4)(Biology). View Reference
      3. Daley SR, Hu DY, Goodnow CC. Helios marks strongly autoreactive CD4+ T cells in two major waves of thymic deletion distinguished by induction of PD-1 or NF-kappaB. J Exp Med. 2013; 210(2):269-285. (Biology). View Reference
      4. Fu W, Ergun A, Lu T, et al. A multiply redundant genetic switch 'locks in' the transcriptional signature of regulatory T cells. Nat Immunol. 2012; 13(10):972-980. (Biology). View Reference
      5. Georgopoulos K. Haematopoietic cell-fate decisions, chromatin regulation and ikaros. Nat Rev Immunol. 2002; 2(3):162-174. (Biology). View Reference
      6. Kelley CM, Ikeda T, Koipally J, et al. Helios, a novel dimerization partner of Ikaros expressed in the earliest hematopoietic progenitors. Curr Biol. 1998; 8(9):508-515. (Biology). View Reference
      7. Serre K, Benezech C, Desanti G, et al. Helios is associated with CD4 T cells differentiating to T helper 2 and follicular helper T cells in vivo independently of Foxp3 expression. PLoS ONE. 2011; 6(6):e20731. (Biology). View Reference
      8. Sugimoto N, Oida T, Hirota K, et al. Foxp3-dependent and -independent molecules specific for CD25+CD4+ natural regulatory T cells revealed by DNA microarray analysis. Int Immunol. 18(8)(Biology). View Reference
      9. Thornton AM, Korty PE, Tran DQ, et al. Expression of Helios, an Ikaros transcription factor family member, differentiates thymic-derived from peripherally induced Foxp3+ T regulatory cells. J Immunol. 2010; 184(7):3433-3441. (Immunogen: ELISA, Flow cytometry, Western blot). View Reference
      10. Yoshimatsu Y, Sujino T, Miyamoto K, et al. Aryl hydrocarbon receptor signals in epithelial cells govern the recruitment and location of Helios+ Tregs in the gut.. Cell Rep. 2022; 39(6):110773. (Clone-specific: Flow cytometry). View Reference
      View All (10) View Less
      571235 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.