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Alexa Fluor® 647 anti-human FOXP3 Antibody

Pricing & Availability
Clone
259D (See other available formats)
Regulatory Status
RUO
Other Names
Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
259D_AF647_FOXP3_Antibody_ICFC_1_041515
Human peripheral blood lymphocytes were surface stained with CD4 Alexa Fluor® 488 and then treated with True-Nuclear™ Transcription Factor Buffer Set. Cells were then stained with FOXP3 (clone 259D) Alexa Fluor® 647 (top) or mouse IgG1, κ Alexa Fluor® 647 isotype control (bottom).
  • 259D_AF647_FOXP3_Antibody_ICFC_1_041515
    Human peripheral blood lymphocytes were surface stained with CD4 Alexa Fluor® 488 and then treated with True-Nuclear™ Transcription Factor Buffer Set. Cells were then stained with FOXP3 (clone 259D) Alexa Fluor® 647 (top) or mouse IgG1, κ Alexa Fluor® 647 isotype control (bottom).
  • 259D_AF647_FOXP3_Antibody_ICFC_2_041515
Compare all formats See Alexa Fluor® 647 spectral data
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320213 25 tests $165
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320214 100 tests $370
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Description

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.

Product Details
Technical data sheet

Product Details

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® 647 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® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 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)
  1. Roncador G, et al. 2005 Eur. J. Immunol. 35:1681.
  2. Yang ZZ, et al. 2006. Blood 107:3639. PubMed
  3. Gavin MA, et al. 2006. P. Natl. Acad. Sci. USA 103:6659. PubMed
  4. Groh V, et al. 2006. Nature Immunology 7:755. PubMed
  5. Tran DQ, et al. 2007. Blood doi:10.1182/blood-2007-06-094656.PubMEd
  6. Long SA, et al. 2008. J Autoimmun. 30:293. PubMed
  7. Gong G, et al. 2009. Blood 113:837. PubMed
  8. Long SA, et al. 2009. Eur J. Immunol. 39:612. PubMed
  9. Long SA, et al. 2010. Diabetes. 59:407. PubMed
  10. Ferraro A, et al. 2014. PNAS. 111:1111. PubMed
  11. Vudattu NK, et al. 2014. J Immunol. 193:587. PubMed
  12. Dupont G, et al. 2014. Cytokine. 69:146. PubMed
Product Citations
  1. VanDyke D, et al. 2022. Cell Rep. 41:111478. PubMed
  2. Krammer S, et al. 2022. Cells. 12: . PubMed
  3. Zhao Q, et al. 2023. Nutrients. 15: . PubMed
  4. Long S, et al. 2009. Eur J Immunol. 39:612. PubMed
  5. Long S, Buckner J 2008. J Autoimmun. 30:293. PubMed
  6. Vudattu N, et al. 2014. J Immunol. 193:587. PubMed
  7. Szabo PA, et al. 2021. Immunity. 54(4):797-814.e6. PubMed
  8. Laing AG, et al. 2020. Nat Med. 26:1623. PubMed
  9. Demaret J, et al. 2015. J Leukoc Biol. 97:791. PubMed
  10. Presicce P, et al. 2011. PLoS One. 6:e28118. PubMed
  11. Ferrer-Font L, et al. 2021. Curr Protoc. 1:e222. PubMed
  12. Gong G, et al. 2009. Blood. 113:837. PubMed
  13. Ibrahim OM, et al. 2021. Cancers (Basel). 13:. PubMed
  14. Leclerc M, et al. 2019. Nat Commun. 10:3345. PubMed
  15. Richardson JR, et al. 2018. Front Immunol. 2.182638889. PubMed
  16. Tran D, et al. 2007. Blood. 110:2983. PubMed
  17. Chen C, et al. 2012. J Immunol. 188:4278. PubMed
  18. Santoni de Sio FR, et al. 2018. J Allergy Clin Immunol. 142:1909. PubMed
  19. Dupont G, et al. 2014. Cytokine. 69:146. PubMed
  20. Ferraro A, et al. 2014. Proc Natl Acad Sci U S A. 111:1111. PubMed
  21. Yang Zhou J, et al. 2022. Front Immunol. 13:898473. PubMed
  22. Mackroth M, et al. 2016. PLoS Pathog. 12:e1005909. PubMed
  23. Long S, et al. 2010. Diabetes. 59:407. PubMed
RRID
AB_492984 (BioLegend Cat. No. 320213)
AB_492984 (BioLegend Cat. No. 320214)

Antigen Details

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 View all products for this Gene ID
UniProt
View information about FOXP3 on UniProt.org

Related FAQs

Can I stain whole blood with anti-FOXP3 using your Foxp3 staining kit?

It is not recommended. It is best to use PBMCs for this testing.

Can FOXP3 be costained with cytokines?

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.

Go To Top Version: 7    Revision Date: 12/13/2016

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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