Alexa Fluor® 488 anti-human Foxp3

RUO: For Research Use Only

human Foxp3 antibody, Alexa Fluor® 488 conjugate

Human PBMCs were surface-stained with PE anti-CD4 (clone RPA-T4) (cat. 12-0049) and APC anti-CD25 (clone BC96) (cat. 17-0259) and subsequently with 20 μl/million cells Alexa 488 anti-human Foxp3 (clone PCH101) or Alexa 488 Rat IgG2a Iso Cntrl using the Foxp3 Buffer Set (cat. 00-5523). The density plot on the left demonstrates co-staining of CD4 and PCH101 while the right plot demonstrates co-staining of CD25 and PCH101. Quadrant lines demarcate the isotype control. Cells in the lymphocyte gate were used for analysis.

Contents: Alexa Fluor® 488 anti-human Foxp3
Catalog Number: 53-4776
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃, gelatin and 0.2% BSA for stabilization
Clone: PCH101
Host/Isotype: Rat IgG2a, κ
Storage Conditions: Store at 4°C.
DO NOT FREEZE.
LIGHT-SENSITIVE MATERIAL.

Product Options
Cat. No.	Size	Price	Add Qty to Cart
53-4776-71	25 tests	To view relevant pricing, please select your country and resume browsing.
53-4776-73	100 tests

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

Research Related Products
Cat. No.	Format
00-5523	eBioscience Foxp3 Staining Buffer Set
12-4777	Phycoerythrin (PE) anti-human Foxp3 (236A/E7)
13-4777	Biotin anti-human Foxp3 (236A/E7)
53-4321	Alexa Fluor® 488 Rat IgG2a Isotype Control
53-4777	Alexa Fluor® 488 anti-human Foxp3 (236A/E7)

Available Formats
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications	Regulatory Status
11-4776	FITC anti-human Foxp3	488	518	IC Flow IH/F IHC(Paraffin)	RUO
12-4776	PE anti-human Foxp3	488	575	IC Flow	RUO
13-4776	Biotin anti-human Foxp3	N/A	N/A	IC Flow IH/F IHC(Paraffin) WB	RUO
14-4776	Affinity Purified anti-human Foxp3	N/A	N/A	IH/F IHC(Paraffin) WB	RUO
15-4776	Phycoerythrin-Cy5 (PE-Cy5) anti-human Foxp3	488	670	IC Flow	RUO
17-4776	APC anti-human Foxp3	633	660	IC Flow	RUO
45-4776	PerCP-Cy5.5 anti-human Foxp3 (PE-Cy5.5 replacement)	488	690	IC Flow	RUO
48-4776	eFluor® 450 anti-human Foxp3	405	450	IC Flow	RUO
51-4776	Alexa Fluor® 647 anti-human Foxp3	633	668	IC Flow	RUO
53-4776	Alexa Fluor® 488 anti-human Foxp3	488	519	IC Flow	RUO
56-4776	Alexa Fluor® 700 anti-human Foxp3 (APC-Cy5.5 Equivalent)	633	723	IC Flow	RUO
57-4776	DISCONTINUED - Pacific Blue® anti-human Foxp3	402	421	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 Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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. 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. This can be used at 20 μl (0.5 μg) per million cells in a 100 μl total staining volume.

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.

Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor Foxp3. Science. 2003 Feb 14;299(5609):1057-61.

Legal

Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc.

Protocol

To view protocol, please download the TDS by clicking the PDF button above.