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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+CD25bright population. The epitope from 236A/E7 is different from that of PCH101 (cat. 72-5776). | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Applications Reported | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
For research use only, not for diagnostic or therapeutic use. 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). This antibody can be used at 5 μl (0.25 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 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] |
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