Exbio — Research products — Antibodies — Nucleus — Anti-Helios Purified

Anti-Helios Purified

Regulatory status

RUO

Antigen

Helios

Clone

22F6

Format

Purified

Reactivity

Mouse, Human

Application

FC (QC tested)

Variant

0.1 mg

11-771-C100

In stock

187.00 USD

0.025 mg

11-771-C025

Delivery 1 week

93.50 USD

Variant

0.1 mg

11-771-C100

In stock

187.00 USD

0.025 mg

11-771-C025

Delivery 1 week

93.50 USD

Contact distributor

Product details

Description

Images

References

SDS download

Isotype

Armenian Hamster IgG

Specificity

The Armenian hamster monoclonal antibody 22F6 recognizes Helios, a transcription factor (intracellular antigen) expressed in some hematopoietic stem cells, and at high levels in thymic-derived regulatory T cells. The epitope is located between amino acids 51 and 107.

Application

FC (QC tested)

Application details

Flow cytometry: Recommended dilution: 1-4 μg/ml.,Recommended protocol: 1) Perform staining of cell surface markers (CD25, CD4 etc.) for 20 min. at room temperature in the dark; 100 μl of peripheral blood. 2) 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. 3) Resuspend the cells in 5 ml of cold fixation solution (Miltenyi Biotec) and incubate for 30 min. on ice. 4) Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. 5) Resuspend the cells in 5 ml of ice-cold PBS with 1% BSA. 6) Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. 7) Resuspend the cells in 5 ml of ice-cold permeabilization solution (Miltenyi Biotec) and incubate 5 min. 8) Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. 9) Resuspend the cells in μl of ice-cold permeabilization solution and add 20 μl of FcR blocking solution and incubate for 5 min. (4°C, in the dark). 10) Perform intracellular staining of Helios for 30 min. (4°C, in the dark) with 10 μl of anti-Helios PE antibody (22F6) in 100 μl. 11) Add 2 ml of ice-cold permeabilization solution and incubate for 5 min. at 4°C in the dark. 12) Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. 13) Resuspend the cells in 3 ml of ice-cold PBS with 1% BSA. 14) Centrifugate for 5 min. at 1000 g, 4°C, and discard the supernatant. 15) 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.

Reactivity

Mouse, Human

Immunogen

Peptide coresponding to the amino acids 51-107 of Helios

Concentration

1 mg/ml

Preparation

Purified by protein-A affinity chromatography.

Formulation

Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide

Storage and handling

Store at 2-8°C. Do not freeze.

The product is intended For Research Use Only. Diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s.

Other names

ANF1A2, IKZF2

Antigen description

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.

Entrez Gene ID 22807

UniProt ID Q9UKS7

Flow cytometry intracellular staining pattern of human peripheral whole blood stained using anti-Helios (22F6) purified antibody (concentration in sample 8 μg/ml, GAM APC).

Flow cytometry multicolor surface staining pattern of human lymphocytes stained using anti-human CD3 (UCHT1) PE-Cy™7 antibody (4 μl reagent / 100 μl of peripheral whole blood) and intracellular staining pattern using anti-Helios (22F6) purified antibody (concentration in sample 8 μg/ml, GAM APC).

Flow cytometry multicolor surface staining pattern of human T cells stained using anti-human CD4 (MEM-241) Pacific Blue™ antibody (4 μl reagent / 100 μl of peripheral whole blood) and intracellular staining pattern using anti-Helios (22F6) purified antibody (concentration in sample 8 μg/ml, GAM APC).

Separation of human CD4 positive Helios positive T cells (red-filled) from CD4 negative Helios negative lymphocytes (black-dashed) in flow cytometry analysis (intracellular staining) of human peripheral whole blood stained using anti-Helios (22F6) purified antibody (concentration in sample 8 μg/ml, GAM APC).

Product specific references:

Serre K, Bénézech 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.
PubMed

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.
PubMed

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.
PubMed

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.
PubMed

Blankenhaus B, Reitz M, Brenz Y, Eschbach ML, Hartmann W, Haben I, Sparwasser T, Huehn J, Kühl 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.
PubMed

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.
PubMed

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.
PubMed

Fourcade J, Sun Z, Chauvin JM, Ka M, Davar D, Pagliano O, Wang H, Saada S, Menna C, Amin R, Sander C, Kirkwood JM, Korman AJ, Zarour HM: CD226 opposes TIGIT to disrupt Tregs in melanoma. JCI Insight. 2018 Jul 26;3(14):e121157.
PubMed

Černý V, Novotná O, Petrásková P Hudcová K, Boráková K, Prokešová L, Kolářová L, Hrdý J: Lower functional and proportional characteristics of cord blood treg of male newborns compared with female newborns. Biomedicines 2021, 9, 170.
PubMed