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Anti-Human Foxp3 Alexa Fluor® 700

Also known as: Forkhead Box P3, Scurfin, JM2, Treg

RUO: For Research Use Only

human Foxp3 antibody, Alexa Fluor® 700 conjugate

Normal human peripheral blood cells were surface stained with Anti-Human CD4 FITC (cat. 11-0049) (left) and Anti-Human CD25 PE (cat. 12-0259) (right) then fixed and permeabilized using the Foxp3 Staining Buffers (cat. 00-5523 and subsequently stained with Anti-Human Foxp3 Alexa Fluor® 700. Cells in the lymphocyte gate were used for analysis.

Contents: Anti-Human Foxp3 Alexa Fluor® 700
Catalog Number: 56-4776
Concentration: 5 μl (0.5 μg)/test
Formulation: aqueous buffer, 0.09% sodium azide, contains stabilizer if necessary
Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material.
Clone: PCH101
Host/Isotype: Rat IgG2a, κ

Product Options
Cat. No.	Size	Price	Add Qty to Cart
56-4776-41	25 tests	To view relevant pricing, please select your country and resume browsing.

Note: Several countries will continue to be supplied via distributors. Country specific prices may apply.

Research Related Products
Cat. No.	Format
00-5523	Foxp3 Staining Buffer Set
14-4774	Anti-Foxp3 Purified (150D/E4)
56-4321	Rat IgG2a K Isotype Control Alexa Fluor® 700

Available Formats
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications	Regulatory Status
11-4776	Anti-Human Foxp3 FITC	488	518	IC Flow IHC (F) IHC (FFPE)	RUO
12-4776	Anti-Human Foxp3 PE	488	575	IC Flow	RUO
13-4776	Anti-Human Foxp3 Biotin	N/A	N/A	IC Flow IHC (F) IHC (FFPE) WB	RUO
14-4776	Anti-Human Foxp3 Purified	N/A	N/A	IHC (F) IHC (FFPE) WB	RUO
15-4776	Anti-Human Foxp3 PE-Cy5	488	670	IC Flow	RUO
17-4776	Anti-Human Foxp3 APC	633	660	IC Flow	RUO
45-4776	Anti-Human Foxp3 PerCP-Cy5.5	488	690	IC Flow	RUO
48-4776	Anti-Human Foxp3 eFluor® 450 (Pacific Blue® replacement)	405	450	IC Flow	RUO
50-4776	Anti-Human Foxp3 eFluor® 660 (Alexa Fluor® 647 Replacement)	633	660	IC Flow	RUO
51-4776	Anti-Human Foxp3 Alexa Fluor® 647 (To Be Discontinued. Refer to NEW Format: eFluor® 660, cat. 50-4776)	633	668	IC Flow	RUO
53-4776	Anti-Human Foxp3 Alexa Fluor® 488	488	519	IC Flow	RUO
56-4776	Anti-Human Foxp3 Alexa Fluor® 700	633	723	IC Flow	RUO
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µL to 5 µL. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.

Description

eBioscience offers a panel of monoclonal antibodies to different epitopes of human Foxp3, providing useful tools for investigating the complete expression pattern of Foxp3 at the protein level, and discerning the precise subsets of Foxp3⁺ cells. Please contact tech@ebioscience.com or 888.810.6168 for any additional assistance.

The PCH101 antibody reacts with the amino terminus of human foxp3 protein also known as FORKHEAD BOX P3, SCURFIN, and JM2; cross reactivity of this antibody to other proteins has not been determined. Foxp3, a 49-55 kDa protein, is a member of the forkhead/winged-helix family of transcriptional regulators, and was identified as the gene defective in ‘scurfy’ (sf) mice. Constitutive high expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg cells), and ectopic expression of foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Intracellular staining of human peripheral blood mononuclear cells (PBMCs) with PCH101 antibody using the anti-human Foxp3 Staining Set and protocol reveals approximately 0.5-4% of lymphocytes staining, with the majority of staining occurring in the CD25^bright population. This is subject to donor variability.

PCH101 crossreacts with rhesus, chimpanzee and cynomolgus. We recommend the use of CD4 (OKT4, cat. 11-0048, or RPA-T4, cat. 11-0049, depending on the species) and CD25 (BC96, cat. 17-0259). Please refer to our crossreactivity table for more information.

Applications Reported

For research use only, not for diagnostic or therapeutic use.

The Alexa Fluor® 700 emits at 723 nm and can be excited with the He-Ne 633 laser. Most instruments will require a 685 LP mirror and 710/20 filter. Please make sure that your instrument is capable of detecting this fluorochome.

This PCH101 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested

This PCH101 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of human peripheral blood leukocytes using the Foxp3 Buffer Set (cat. 00-5523) and protocol. Please click here for Staining Protocol (refer to Protocol B: One-step protocol for intracellular (nuclear) proteins). This antibody can be used at 5 μl (0.5 μg)/per test. A test is defined as the amount (μg)/test of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10⁵ to 10⁸ cells/test.

Special Notes

Please see the following link for FAQ regarding the usage of eBioscience Foxp3 reagents:
http://www.ebioscience.com/ebioscience/Foxp3FAQs.htm

The staining protocol has been optimized with Ficoll prepped PBMCs. The use of lysed whole blood is not recommended.

It is critical that this antibody be used in conjunction with the Foxp3 Staining Buffers (cat 00-5523) for flow cytometric analysis.

References

Bell MP, Svingen PA, Rahman MK, Xiong Y, Faubion WA Jr. FOXP3 regulates TLR10 expression in human T regulatory cells. J Immunol. 2007 Aug 1;179(3):1893-900. (PCH101, ChIP assay, PubMed)

Clark RA, Kupper TS. IL-15 and dermal fibroblasts induce proliferation of natural regulatory cells isolated from human skin. Blood. 2007 Jan 1;109(1):194-202.(PCH101, IHC frozen, PubMed)

Epple HJ, Loddenkemper C, Kunkel D, Tröger H, Maul J, Moos V, Berg E, Ullrich R, Schulzke JD, Stein H, Duchmann R, Zeitz M, Schneider T. Mucosal but not peripheral FOXP3+ regulatory T cells are highly increased in untreated HIV infection and normalize after suppressive HAART. Blood. 2006 Nov 1;108(9):3072-8. PCH101, IHC paraffin, PubMed)

Manigold T, Shin EC, Mizukoshi E, Mihalik K, Murthy KK, Rice CM, Piccirillo CA, Rehermann B. Foxp3+CD4+CD25+ T cells control virus-specific memory T cells in chimpanzees recovered from Hepatitis C.Blood. 2006 Jun 1;107(11):4424-32. (PCH101 in chimpanzee, PubMed)

Ahmadzadeh M, Rosenberg SA. IL-2 Administration Increases CD4+CD25hiFoxp3+ Regulatory T Cells in Cancer Patients. Blood. 2006 Mar 15;107(6):2409-14. (PCH101, IC Flow, PubMed)

Hartwig UF, Nonn M, Khan S, Meyer RG, Huber C, Herr W. Depletion of alloreactive T cells via CD69: implications on antiviral, antileukemic and immunoregulatory T lymphocytes. Bone Marrow Transplant. 2006 Feb;37(3):297-305. (PCH101, IC Flow, PubMed)

Crellin NK, Garcia RV, Hadisfar O, Allan SE, Steiner TS, Levings MK. Human CD4+ T Cells Express TLR5 and Its Ligand Flagellin Enhances the Suppressive Capacity and Expression of FOXP3 in CD4+CD25+ T Regulatory Cells. J Immunol. 2005 Dec 15;175(12):8051-9. (PCH101, IC Flow, PubMed)

Lim, H.W., P. Hillsamer, A.H. Banham, and C.H. Kim. 2005. Cutting Edge: Direct Suppression of B cells by CD4+CD25+ Regulatory T cells. J Immunol. 2005 Oct 1;175(7):4180-3. (PCH101,IC Flow, PubMed)

Manavalan JS, Kim-Schulze S, Scotto L, Naiyer AJ, Vlad G, Colombo PC, Marboe C, Mancini D, Cortesini R, Suciu-Foca N. Alloantigen specific CD8+CD28- FOXP3+ T suppressor cells induce ILT3+ ILT4+ tolerogenic endothelial cells, inhibiting alloreactivity. Int Immunol. 2004 Aug;16(8):1055-68.

Takahata Y, Nomura A, Takada H, Ohga S, Furuno K, Hikino S, Nakayama H, Sakaguchi S, Hara T. CD25+CD4+ T cells in human cord blood: an immunoregulatory subset with naïve phenotype and specific expression of forkhead box p3 (Foxp3) gene. Exp Hematol. 2004 Jul;32(7):622-9.

Sakaguchi S. 2003. The origin of FOXP3-expressing CD4+ regulatory T cells: thymus or periphery. J Clin Invest. 2003 November 1; 112(9): 1310–1312.

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