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.

PRODUCT DETAILS

Clone: 22F6

Specificity: Helios

Reactivity: Human|Mouse

Format: Purified

Isotype: Armenian Hamster IgG

Application: FC-IC

Storage: 2-8°C protected from light.Do not freeze

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

Data Sheets: TDS | SDS

RECOMMENDED USAGE

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.

REFERENCES

1. Serre K, et al. 2011. PLoS One 6:e20731.
2. Thornton AM, et al. 2010. J Immunol 184:3433-41.
3. McIver Z, et al. 2013. Haematologica 98:346-52.
4. Atarashi K, et al. 2013. Nature 500:232-6.
5. Blankenhaus B, et al. 2014. PLoS Pathog 10:e1003913.
6. Daley SR, et al. 2013. J Exp Med 210:269-85.
7. Pinheiro D, et al. 2011. Immunology 132:111-22.
8. Fourcade J, et al. 2018. JCI Insight 3.
9. Černý V, et al. 2021. Biomedicines 9.

For research use only. Not intended for use in diagnostic procedures.