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Alexa Fluor® 488 Armenian Hamster Anti-Helios
Alexa Fluor® 488 Armenian Hamster Anti-Helios
Two-color flow cytometric analysis of Helios expression Left Panel: Human peripheral blood mononuclear cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with Alexa Fluor® 647 Mouse Anti-Human CD4 antibody (Cat No. 557707) and either Alexa Fluor® 488 Hamster IgG2, κ Isotype Control (Cat No. 562168, Left Plot) or Alexa Fluor® 488 Armenian Hamster Anti-Helios antibody (Cat. No. 563950; Right Plot). Right Panel: Mouse splenic leucocytes were similarly fixed and permeabilized with the BD Transcription Factor Buffer Set. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat No. 553730/557308/561829) and either Alexa Fluor® 488 Hamster IgG2, κ Isotype Control (Left Plot) or Alexa Fluor® 488 Armenian Hamster Anti-Helios antibody (Right Plot). The two-color flow cytometric contour plots showing the correlated expression patterns 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 cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of Helios expression Left Panel: Human peripheral blood mononuclear cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with Alexa Fluor® 647 Mouse Anti-Human CD4 antibody (Cat No. 557707) and either Alexa Fluor® 488 Hamster IgG2, κ Isotype Control (Cat No. 562168, Left Plot) or Alexa Fluor® 488 Armenian Hamster Anti-Helios antibody (Cat. No. 563950; Right Plot). Right Panel: Mouse splenic leucocytes were similarly fixed and permeabilized with the BD Transcription Factor Buffer Set. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat No. 553730/557308/561829) and either Alexa Fluor® 488 Hamster IgG2, κ Isotype Control (Left Plot) or Alexa Fluor® 488 Armenian Hamster Anti-Helios antibody (Right Plot). The two-color flow cytometric contour plots showing the correlated expression patterns 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 cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
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
AB_2738505
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 Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563950 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.

  

563950 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
563950 Rev.1
Citations & References
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View product citations for antibody "563950" on CiteAb

Development References (9)

  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
View All (9) View Less
563950 Rev. 1

 

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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.