Biolegend, 320212, Alexa Fluor® 488 anti-human FOXP3 Antibody, 100tests

FOXP3 is a 50-55 kD transcription factor, also known as Forkhead box protein P3, Scurfin, JM2, or IPEX. It is proposed to be a master regulatory gene and more specific marker of T regulatory cells than most cell surface markers (such as CD4 and CD25). Transduced expression of FOXP3 in CD4 + /CD25 - cells has been shown to induce GITR, CD103, and CTLA4 and impart a T regulatory cell phenotype. FOXP3 is mutated in X-linked autoimmunity-allergic dysregulation syndrome (XLAAD or IPEX) in humans and in "scurfy" mice. Overexpression of FOXP3 has been shown to lead to a hypoactive immune state suggesting that this transcriptional factor is a central regulator of T cell activity. In human, unlike in mouse, two isoforms of FOXP3 have been reported: one (FOXP3) corresponding to the canonical full-length sequence; the other (FOXP3 δ2) lacking exon 2. The 259D antibody recognizes human FOXP3 epitope in the region of amino acids 105-235.
100tests
Verified Reactivity: Human
Reported Reactivity: Cynomolgus, Rhesus, Baboon, Chimpanzee
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Full-length FOXP3 protein
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
Concentration: Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling: The FOXP3 antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: ICFC - Quality tested
Recommended Usage: Each lot of this antibody is quality control tested by intracellular flow cytometry using our True-Nuclear™ Transcription Factor Staining Protocol. For flow cytometric staining, the suggested use of this reagent is 5 µl per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses
Excitation Laser: Blue Laser (488 nm)
Application Notes: Additional reported applications (for the relevant formats) include: Western blotting1, and immunohistochemical staining1 of acetone-fixed frozen sections and formalin-fixed paraffin-embedded sections. The 259D antibody gives strong positivity on paraffin and frozen sections and the antibody stains some epithelial cells. The binding of 206D to FOXP3 can be partially blocked by 259D, but 206D does not show significant blocking effect on 259D binding. NOTE: For flow cytometric staining with this clone, True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) offers improved staining and is highly recommended.
Application References(PubMed link indicates BioLegend citation): Roncador G, et al. 2005 Eur. J. Immunol. 35:1681. Yang ZZ, et al. 2006. Blood 107:3639. PubMed Gavin MA, et al. 2006. P. Natl. Acad. Sci. USA 103:6659. PubMed Groh V, et al. 2006. Nature Immunology 7:755. PubMed Tran DQ, et al. 2007. Blood doi:10.1182/blood-2007-06-094656.PubMEd Long SA, et al. 2008. J Autoimmun. 30:293. PubMed Gong G, et al. 2009. Blood 113:837. PubMed Long SA, et al. 2009. Eur J. Immunol. 39:612. PubMed Long SA, et al. 2010. Diabetes. 59:407. PubMed Ferraro A, et al. 2014. PNAS. 111:1111. PubMed Vudattu NK, et al. 2014. J Immunol. 193:587. PubMed Dupont G, et al. 2014. Cytokine. 69:146. PubMed
Product Citations: Geller M, et al. 2011. Cytotherapy. 13:98. PubMed Huijts C, et al. 2016. Clin Immunol. 168:47-54. PubMed Rech A, et al. 2012. Sci Transl Med. 4:134ra62. PubMed Ritacco C, et al. 2023. iScience. 26:106085. PubMed Ke F, et al. 2023. Elife. 12:. PubMed Hu M, et al. 2022. Int J Mol Sci. 23: . PubMed Kim CJ, et al. 2018. Immunity. 49:1034. PubMed Rapoport A, et al. 2011. Blood. 117:788. PubMed Bamidele AO, et al. 2018. Cell Mol Gastroenterol Hepatol. 7:55. PubMed Sutavani R, et al. 2013. J Immunol. 191:5895. PubMed Kobayashi Y, et al. 2020. Int J Oncol. 999:56. PubMed Huijts CM, et al. 2019. Cancer Immunol Immunother. 68:503. PubMed Detry O, et al. 2017. J Hepatol. 10.1016/j.jhep.2017.03.001. PubMed Giovannetti A, et al. 2013. PLoS One. 18:74332. PubMed Valle A, et al. 2015. J Immunol. 194:2117. PubMed Boularan C, et al. 2015. J Immunol. 195: 2090-2102. PubMed Fox B, et al. 2008. Blood. 111:3897. PubMed Ritacco C, et al. 2021. Bone Marrow Transplant. 56:1828. PubMed He T, et al. 2015. J Virol. 89: 9616-9630. PubMed Cassani B, et al. 2010. J Allergy Clin Immunol. 125:209. PubMed Gavin M, et al. 2006. Proc Natl Acad Sci U S A. 103:6659. PubMed Keefe RC, et al. 2021. Sci Rep. 11:14933. PubMed Monti P, et al. 2008. Diabetes. 57:2341. PubMed Hu M, et al. 2019. Nat Commun. 10:3031. PubMed Sivanandham R, et al. 2020. J Virol. 94. PubMed Farhat AM, et al. 2021. Cell Reports. 35(4):109044. PubMed Tresoldi E, et al. 2011. Haematologica. 96:1357. PubMed Vonderheide R, et al. 2010. Clin Cancer Res. 3.086805556. PubMed Torgerson T, et al. 2009. J Immunol. 183:907. PubMed Faustman D, et al. 2012. PLoS One. 7:e41756. PubMed
RRID: AB_430886 (BioLegend Cat. No. 320211) AB_430887 (BioLegend Cat. No. 320212)
Structure: Forkhead/winged-helix transcription factor family, approximately 50 kD, contains zinc finger and forkhead domains
Distribution: Nuclear; expressed in T regulatory cells
Function: Transcription factor proposed to be a master regulatory gene in T regulatory cell development and a critical factor for immune homeostasis
Interaction: Interacts with DNA
Cell Type: Tregs
Biology Area: Cell Biology, Immunology, Transcription Factors
Molecular Family: Nuclear Markers
Antigen References: 1. Hori S, et al. 2003. Science 299:1057.
Regulation: FOXP3 is present at high levels in T regulatory cell can also be induced by T cell activation
Gene ID: 50943
UniProt: View information about FOXP3 on UniProt.org
Clone: 259D
Regulatory Status: RUO
Other Names: Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
Isotype: Mouse IgG1, κ
Q: Can I stain whole blood with anti-FOXP3 using your Foxp3 staining kit?
A: It is not recommended. It is best to use PBMCs for this testing.
Q: Can FOXP3 be costained with cytokines?
A: The larger holes created by the nuclear permeabilization required for FOXP3 may allow cytokines to leak out of the cell, making it harder to detect lowly-expressed cytokines. You may have to use a control where the cells are only permeabilized through the cell membrane.