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True-Nuclear™ One Step Staining Human Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP)

Pricing & Availability
Clone
206D (See other available formats)
Regulatory Status
RUO
Other Names
Forkhead box protein P3, Scurfin, JM2, IPEX, Zinc finger protein JM2
Isotype
Mouse IgG1, κ
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Product Citations
publications
1_206D_True-Nuclear_AF488_1SKit_060915
Human peripheral blood lymphocytes were stained with True-Nuclear™ One Step Staining Human Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP).
  • 1_206D_True-Nuclear_AF488_1SKit_060915
    Human peripheral blood lymphocytes were stained with True-Nuclear™ One Step Staining Human Treg Flow™ Kit (FOXP3 Alexa Fluor® 488/CD25 PE/CD4 PerCP).
  • 2_206D_True-Nuclear_AF488_1SKit_060915
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Cat # Size Price Save
320127 25 tests ¥128,920
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 206D 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
Baboon, Cynomolgus, Rhesus, Pigtailed Macaque
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Full-length FOXP3 protein
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
This kit is guaranteed for six months. Upon receipt, store between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICFC - Quality tested
Recommended Usage

Materials Provided:
1. Alexa Fluor® 488 anti-human FOXP3/CD25 PE/CD4 PerCP antibody cocktail - 25 tests
2. Alexa Fluor® 488 Mouse IgG1, κ isotype control/CD25 PE/CD4 PerCP antibody cocktail - 25 tests
3. True-Nuclear™ Transcription Buffer Set - 120 tests (Cat. No. 424401)

Materials not included:
1. Cell Staining Buffer (Cat. No. 420201)
2. Single Color Compensation Controls

Immunofluorescence Staining Procedures:

1. Aliquot 100 µL of target cells to each tube.
2. Add 1 mL of the Transcription Factor 1X Fix solution to each tube, vortex, and incubate at room temperature in the dark for 30-60 minutes.
3. Without washing, add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
4. Centifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
5. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
6. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
7. Resuspend the cell pellet in 100 µL of the Transcription Factor 1X Perm Buffer.
8. Add 20 µL of Alexa Fluor® 488 anti-human FOXP3/CD25 PE/CD4 PerCP antibody cocktail or 20 µL of Alexa Fluor® 488 mouse IgG1, κ isotype control/CD25 PE/CD4 PerCP antibody cocktail into the appropriate tubes. Incubate in the dark at room temperature for at least 30 minutes.
9. Add 2 mL of the Transcription Factor 1X Perm Buffer to each tube.
10. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
11. Add 2 mL of the cell staining buffer.
12. Centrifuge tubes at 400 x g at room temperature for five minutes, and discard the supernatant.
13. Resuspend in 0.5 mL cell staining buffer and then acquire tubes on a flow cytometer.

Caution: The True-Nuclear™ Transcription Factor Buffer Set contains paraformaldehyde, which is toxic and mutagenic. Please handle with caution. Wear gloves, lab coats, and necessary protection to avoid direct contact.

NOTE: For flow cytometric staining with this clone, True-Nuclear™ Transcription Factor Buffer Set (Cat. No. 424401) offers improved staining and is highly recommended over the Foxp3/Perm Buffer Set (Cat. No. 421403) and the Nuclear Factor Fixation and Permeabilization Buffer Set (Cat. No. 422601).

Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections1 and formalin-fixed paraffin-embedded sections1, 8, 19-20, and Western blotting1. 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. (IHC)
  2. Yang ZZ, et al. 2006. Blood 107:3639.
  3. Liu W, et al. 2006. J. Exp. Med. 203:1701.PubMed
  4. Bollyky PL, et al. 2007. J. Immunol. 179:744.
  5. Bell MP, et al. 2007. J. Immunol. 179:1893.
  6. Tran DQ, et al. 2007. Blood doi:10.1182/blood-2007-06-094656. PubMed
  7. Gao Q,et al.2007.J Clin Oncol.25:2586.(IHC) PubMed
  8. Pillai V,et al. 2008. Blood 111:463.PubMed
  9. Zheng Y, et al. 2008. J. Immunol. 181:1683. PubMed
  10. Zonios DI, et al. 2008. Blood112:287. PubMed
  11. Kavanagh B, et al. 2008. Blood. PubMed
  12. Nevala WK, et al. 2009. Clin Cancer Res. 15:1931. PubMed
  13. Grant J, et al. 2009. Cytometry B Clin Cytom. 76:69. PubMed
  14. Nigam P, et al. 2010. J. Immunol. 184:1690. PubMed
  15. Kmieciak M, et al. 2009. J. Transl. Med. 7:89. (ICFC) PubMed
  16. Hartigan-O'Connor DJ,et al. 2007. J Exp Med.204:2679. PubMed
  17. Raghaven S, et al. 2009. Ann Rheum Dis. 68:1908. PubMed
  18. Hodi FS, et al. 2014. Cancer Immunol Res. 2:632.(IHC) PubMed
  19. Sziros E, et al. 2015. Clin Cancer Res. 21:2840.(IHC) PubMed
Product Citations
  1. Park S, et al. 2016. Proc Natl Acad Sci U S A. 113(52):E8379-E8386. PubMed

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
CD Molecules, Nuclear Markers
Antigen References

1. Hori S, et al. 2003. Science 299:1057.
2. Gandhi R, et al. 2010. Nat. Immunol. 11:846.

Regulation
FOXP3 is present at high levels in T regulatory cells, it can also be induced by T cell activation.
Gene ID
50943 View all products for this Gene ID 50943 View all products for this Gene ID 20371 View all products for this Gene ID 317382 View all products for this Gene ID 916 View all products for this Gene ID
UniProt
View information about FOXP3 on UniProt.org

Related FAQs

How stable is PerCP/Cy5.5 tandem as compared to PerCP alone?

PerCP/Cy5.5 is quite photostable and also better than PerCP alone in withstanding fixation.

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: 3    Revision Date: 12/16/2021

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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