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Anti-Human Foxp3 Alexa Fluor® 488

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

RUO: For Research Use Only

Contents: Anti-Human Foxp3 Alexa Fluor® 488
Catalog Number: 53-4777
Concentration: Suffix -71/73, 20 μL (0.5 μg)/test; Suffix -41/42, 5 μL (0.5 μg)/test
Formulation: aqueous buffer, 0.09% sodium azide, may contain carrier protein/stabilizer
Storage Conditions: Store at 2-8°C. Do not freeze. Light sensitive material.
Clone: 236A/E7
Host/Isotype: Mouse IgG1

Product Options
Cat. No.	Size	Price	Add Qty to Cart
53-4777-71	25 tests	To view relevant pricing, please select your country and resume browsing.
53-4777-73	100 tests

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

Research Related Products
Cat. No.	Format
00-5521	Foxp3 Fixation/Permeabilization Concentrate and Diluent
00-5523	Foxp3 Staining Buffer Set
12-0259	Anti-Human CD25 PE (BC96)
53-4714	Mouse IgG1 K Isotype Control Alexa Fluor® 488

Available Formats
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications	Regulatory Status
11-4777	Anti-Human Foxp3 FITC	488	518	IC Flow	RUO
12-4777	Anti-Human Foxp3 PE	488	575	IC Flow	RUO
13-4777	Anti-Human Foxp3 Biotin	N/A	N/A	IC Flow IHC (F) IHC (FFPE)	RUO
14-4777	Anti-Human Foxp3 Purified	N/A	N/A	IC Flow IHC (F) IHC (FFPE) WB	RUO
17-4777	Anti-Human Foxp3 APC	633	660	IC Flow	RUO
48-4777	Anti-Human Foxp3 eFluor® 450 (Pacific Blue® replacement)	405	450	FC	RUO
51-4777	Anti-Human Foxp3 Alexa Fluor® 647	633	668	IC Flow	RUO
53-4777	Anti-Human Foxp3 Alexa Fluor® 488	488	519	IC Flow IHC (F) IHC (FFPE)	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. Other antibodies to human Foxp3 include the recently published PCH101 (cat. 72-5776) and ebio7979 (cat. 14-7979).

The 236A/E7 antibody reacts with 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 and flow cytometric analysis of freshly isolated human peripheral blood mononuclear cells (PBMCs) with the 236A/E7 antibody using the Foxp3 Staining Buffer Set (cat. 00-5523) and protocol reveals staining of the CD4+CD25^bright population.

The epitope from 236A/E7 is different from that of PCH101 (cat. 72-5776).

Applications Reported

This 236A/E7 antibody has been reported for use in intracellular staining followed by flow cytometric analysis and IHC.

Applications Tested

This 236A/E7 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of normal human peripheral blood mononuclear cells 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). Refer to catalog number suffix on the vial for amount to use per test: 71/73 are 20 μL (0.5 μg) per test; whereas 41/42 are 5 μL (0.5 μg) per test. A test is defined as the amount (μg) 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 freshly 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

Sakaguchi S. 2003. The origin of FOXP3-expressing CD4+ regulatory T cells: thymus or periphery. J Clin Invest. 112(9):1310-2.

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

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

Alvaro T, Lejeune M, Salvado MT, Bosch R, Garcia JF, Jaen J, Banham AH, Roncador G, Montalban C, Piris MA. 2005. Outcome in Hodgkin's lymphoma can be predicted from the presence of accompanying cytotoxic and regulatory T cells. Clin Cancer Res. 11(4):1467-73. [IHC paraffin using 236A/E7, 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. 175: 4180-4183. (236A/E7, IH(frozen), PCH101, IC (flow), PubMed)

Roncador, G., P.J. Brown, L. Maestre, S. Hue, J.L. Martinez-Torrecuadrada, K.L. Ling, S. Pratap, C. Toms, B.C. Fox, V. Cerundolo, F. Powrie, and A.H. Banham. 2005. Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level. Eur J Immunol. 35: 1681-91. [236A/E7, IC(flow), IH(paraffin), PubMed]

Roncador G, Garcia JF, Garcia JF, Maestre L, Lucas E, Menarguez J, Ohshima K, Nakamura S, Banham AH, Piris MA. 2005. FOXP3, a selective marker for a subset of adult T-cell leukaemia/lymphoma. Leukemia [Epub ahead of print] [IHC paraffin sections using 236A/E7, PubMed]

Wolf D, Wolf AM, Rumpold H, Fiegl H, Zeimet AG, Muller-Holzner E, Deibl M, Gastl G, Gunsilius E, Marth C.The expression of the regulatory T cell-specific forkhead box transcription factor FoxP3 is associated with poor prognosis in ovarian cancer. Clin Cancer Res. 2005 Dec 1;11(23):8326-31. [IHC paraffin using 236A/E7, PubMed]

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.

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