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.
Kreuzreaktivität (Details)
Human, Mouse
Aufreinigung
Purified by protein-A affinity chromatography.
Reinheit
> 95 % (by SDS-PAGE)
Immunogen
Peptide coresponding to the amino acids 51-107 of Helios
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.
Beschränkungen
Nur für Forschungszwecke einsetzbar
Konzentration
1 mg/mL
Buffer
Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide
Konservierungsmittel
Sodium azide
Vorsichtsmaßnahmen
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Lagerung
4 °C
Informationen zur Lagerung
Store at 2-8°C. Do not freeze.
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Serre, Bénézech, Desanti, Bobat, Toellner, Bird, Chan, Kastner, Cunningham, Maclennan, Mohr: "Helios is associated with CD4 T cells differentiating to T helper 2 and follicular helper T cells in vivo independently of Foxp3 expression." in: PLoS ONE, Vol. 6, Issue 6, pp. e20731, (2011) (PubMed).
Pinheiro, Singh, Grant, Appleton, Sacchini, Walker, Chadbourne, Palmer, Armitage-Chan, Thompson, Williamson, Cunningham, Garden: "Phenotypic and functional characterization of a CD4(+) CD25(high) FOXP3(high) regulatory T-cell population in the dog." in: Immunology, Vol. 132, Issue 1, pp. 111-22, (2010) (PubMed).
IKAROS family zinc finger 2,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.,ANF1A2, IKZF2