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Hamster Monoclonal to Helios

22F6 (IgG)

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Background

Helios, also known as IKZF2 (Ikaros family zinc finger protein 2) is a hematopoietic-specific transcription factor involved in the regulation of lymphocyte development, together with other members of this family, such as Aiolos and Ikaros. Helios forms homo- and heterodimers with these proteins and is thought to function predominantly in early hematopoietic development. Expression of Helios, Aiolos and Ikaros is restricted to cells of the hematopoietic system, whereas other family members, Eos and Pegassus, are more widely expressed. Helios is expressed at early stages of thymocyte development. In mature T cells, Helios has been strongly associated with Treg cells.

Specificity

The armenian hamster monoclonal antibody 22F6 recognizes Helios, a transcription factor expressed in some hematopoietic stem cells, and at high levels in thymic-derived regulatory T cells.

Regulatory Status

Immunogen

Peptide coresponding to the amino acids 51-107 of Helios

Species Reactivity:

  • Human
  • Mouse

Negative Species:

Applications:

  • Flow Cytometry
    Staining method:
    - Perform staining of cell surface markers (CD25, CD4 etc.) for 20 min. at room temperature in the dark; 100 µl of peripheral blood.
    - Add 3 ml of PBS with 1% BSA, centrifugate at 300g and discard the supernatant. Further steps perform on ice and with ice-cold reagents.
    - Resuspend the cells in 5 ml of cold fixation solution (Miltenyi Biotec) and incubate for 30 min. on ice.
    - Centrifugate for 5 min. at 1000 g, 4C, and discard the supernatant.
    - Resuspend the cells in 5 ml of ice-cold PBS with 1% BSA.
    - Centrifugate for 5 min. at 1000 g, 4C, and discard the supernatant.
    - Resuspend the cells in 5 ml of ice-cold permeabilization solution (Miltenyi Biotec) and incubate 5 min.
    - Centrifugate for 5 min. at 1000 g, 4C, and discard the supernatant.
    - Resuspend the cells in 80µl of ice-cold permeabilization solution and add 20 µl of FcR blocking solution and incubate for 5 min. (4C, in the dark).
    - Perform intracellular staining of Helios for 30 min. (4C, in the dark) with 10 µl of anti-Helios PE antibody (22F6) in 100 µl.
    - Add 2 ml of ice-cold permeabilization solution and incubate for 5 min. at 4C in the dark.
    - Centrifugate for 5 min. at 1000 g, 4C, and discard the supernatant.
    - Resuspend the cells in 3 ml of ice-cold PBS with 1% BSA.
    - Centrifugate for 5 min. at 1000 g, 4C, and discard the supernatant.
    - Resuspend the cells in 150 µl of ice-cold PBS with 1% BSA and measure on a flow cytometry device with appropriate setting as soon as possible. Keep cold until measuring.
Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

Product Specific References

  • *McIver Z, Melenhorst JJ, Wu C, Grim A, Ito S, Cho I, Hensel N, Battiwalla M, Barrett AJ: Donor lymphocyte count and thymic activity predict lymphocyte recovery and outcomes after matched-sibling hematopoietic stem cell transplant. Haematologica. 2013 Mar;98(3):346-52. [Abstract]
  • *Atarashi K, Tanoue T, Oshima K, Suda W, Nagano Y, Nishikawa H, Fukuda S, Saito T, Narushima S, Hase K, Kim S, Fritz JV, Wilmes P, Ueha S, Matsushima K, Ohno H, Olle B, Sakaguchi S, Taniguchi T, Morita H, Hattori M, Honda K: Treg induction by a rationally selected mixture of Clostridia strains from the human microbiota. Nature. 2013 Aug 8;500(7461):232-6. [Abstract]
  • *Blankenhaus B, Reitz M, Brenz Y, Eschbach ML, Hartmann W, Haben I, Sparwasser T, Huehn J, Khl A, Feyerabend TB, Rodewald HR, Breloer M. Foxp3⁺ regulatory T cells delay expulsion of intestinal nematodes by suppression of IL-9-driven mast cell activation in BALB/c but not in C57BL/6 mice. PLoS Pathog. 2014 Feb 6;10(2):e1003913. [Abstract] [Full Text]
  • *Daley SR1, 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-κB. J Exp Med. 2013 Feb 11;210(2):269-85. [Abstract] [Full Text]
  • *Pinheiro D, Singh Y, Grant CR, Appleton RC, Sacchini F, Walker KR, Chadbourne AH, Palmer CA, Armitage-Chan E, Thompson I, Williamson L, Cunningham F, Garden OA. Phenotypic and functional characterization of a CD4(+) CD25(high) FOXP3(high) regulatory T-cell population in the dog. Immunology. 2011 Jan;132(1):111-22. [Abstract] [Full Text]
  • *Serre K, Bnzech C, Desanti G, Bobat S, Toellner KM, Bird R, Chan S, Kastner P, Cunningham AF, Maclennan IC, Mohr E: 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. [Abstract]
  • *Thornton AM, Korty PE, Tran DQ, Wohlfert EA, Murray PE, Belkaid Y, Shevach EM: Expression of Helios, an Ikaros transcription factor family member, differentiates thymic-derived from peripherally induced Foxp3+ T regulatory cells. J Immunol. 2010 Apr 1;184(7):3433-41. [Abstract]
  • For research use only. Not for drug, diagnostic or other use.

    Example Data

    Helios
    Fig. 1


    Fig. 1. Human peripheral blood lymphocytes were stained with anti-CD4 Pacific BlueTM, fixed and permeabilized with Treg Detection Kit (Miltenyi cat. 130-094-163), and then intracellulary stained with anti-Helios PE (clone 22F6).


    22F6 PE-Cy7


    Fig. 2. Intracellular staining of CD4+ human peripheral blood lymphocytes using anti-Helios (clone 22F6) PE-CyTM7.

    Unless indicated otherwise, all products are For Research Use Only and not for diagnostic or therapeutic use. Not for resale or transfer either as a stand-alone product or as a component of another product without written consent of EXBIO. EXBIO will not be held responsible for patent infringement or other violations that may occur with the use of our products. All orders are accepted subject to EXBIOs term and conditions which are available at www.exbio.cz.
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