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RY586 Armenian Hamster Anti-Helios
RY586 Armenian Hamster Anti-Helios
Two-color flow cytometric analysis of Helios expression    Left Panel: Human peripheral blood mononuclear cells (PBMC) were stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425) 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™ RY586 Armenian Hamster IgG2 κ, Isotype Control (Cat. No. 568823; Left Plot) or BD Horizon™ RY586 Armenian Hamster Anti-Helios antibody (Cat. No. 568526/568527; Right Plot).    Right Panel: Mouse splenic leucocytes were stained with BD OptiBuild™ BV421 Rat Anti-Mouse CD4 antibody (Cat. No. 740007) and fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set. The cells were then stained with either Armenian Hamster IgG2 κ, Isotype Control (Left Plot) or BD Horizon™ RY586 Armenian Hamster Anti-Helios antibody (Right Plot). The 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 leucocytes.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Two-color flow cytometric analysis of Helios expression    Left Panel: Human peripheral blood mononuclear cells (PBMC) were stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425) 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™ RY586 Armenian Hamster IgG2 κ, Isotype Control (Cat. No. 568823; Left Plot) or BD Horizon™ RY586 Armenian Hamster Anti-Helios antibody (Cat. No. 568526/568527; Right Plot).    Right Panel: Mouse splenic leucocytes were stained with BD OptiBuild™ BV421 Rat Anti-Mouse CD4 antibody (Cat. No. 740007) and fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set. The cells were then stained with either Armenian Hamster IgG2 κ, Isotype Control (Left Plot) or BD Horizon™ RY586 Armenian Hamster Anti-Helios antibody (Right Plot). The 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 leucocytes.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
商品详情
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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)
5 µl
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


准备和存储

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 unreacted dye was removed.

推荐的实验流程

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.

商品通知

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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. CF™ is a trademark of Biotium, Inc.
抗体详情
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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.

格式详情
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RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 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), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RY586
Yellow-Green 561 nm
564 nm
586 nm
报价单和参考
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View product citations for antibody "568527" on CiteAb

研发参考 (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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
  5. Georgopoulos K. Haematopoietic cell-fate decisions, chromatin regulation and ikaros. Nat Rev Immunol. 2002; 2(3):162-174. (Biology). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
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