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![R718 Mouse Anti-Human FoxP3](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/566936_base/566936Image1.png)
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![R718 Mouse Anti-Human FoxP3](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/566936_base/566936Image1.png)
Multicolor flow cytometric analysis of FoxP3 expressed in resting human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stained with FITC Mouse Anti-Human CD4 (Cat. No. 566320) and BD Horizon™ BV421 Mouse Anti-Human CD25 (Cat. No. 562442) antibodies. The cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) followed by intracellular staining with BD Horizon™ R718 Mouse Anti-Human FoxP3 antibody (Cat No. 566935/566936/566937). The pseudocolor density plots showing the corelated expression of FoxP3 versus CD25 (Left Plot) or CD4 versus FoxP3 (Right Plot) were derived from gated events with the forward and side-light scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
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![R718 Mouse Anti-Human FoxP3](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/566936_base/566936Image1.png)
BD Horizon™ R718 Mouse Anti-Human FoxP3
![R718 Mouse Anti-Human FoxP3](/content/dam/bdb/products/global/reagents/flow-cytometry-reagents/research-reagents/single-color-antibodies-ruo/566936_base/566937_01.png)
R718 Mouse Anti-Human FoxP3
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Preparation And Storage
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
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- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
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The 259D/C7 monoclonal antibody specifically recognizes human Forkhead box protein P3 (FoxP3) that is also known as Scurfin. FoxP3 is encoded by FOXP3 (Forkhead box P3), likewise known as IPEX (Immune Dysregulation, Polyendocrinopathy, Enteropathy, X-Linked) and JM2, that belongs to the forkhead/winged-helix family of transcriptional regulators. FoxP3 is expressed in CD4+ regulatory T cells (Treg) and represents a specific marker for these cells. Flow cytometric analyses have shown that FoxP3 is expressed by the majority of CD4+CD25+high T cells in peripheral blood while less than half of the CD4+CD25+intermediate cell population are FoxP3 positive. Approximately 5-10% of peripheral CD4+ cells are CD4+CD25+ T regulatory cells. T regulatory cells are thought to play a critical role in the regulation of T cell-mediated immunity and to protect against autoimmunity by suppressing the proliferation and cytokine production of other T cells. In support of this hypothesis, it has been found that Foxp3 is mutated in Scurfy (sf) mice that have defective T cell tolerance leading to an X-linked lymphoproliferative disease. The 259D/C7 antibody recognizes all currently-identified isoforms of human FoxP3 and is crossreactive with FoxP3 from Cynomolgus, Rhesus and Baboon primates.
The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor® 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor® 700.
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Development References (4)
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Brunkow ME, Jeffery EW, Hjerrild KA, et al. Disruption of a new forkhead/winged-helix protein, scurfin, results in the fatal lymphoproliferative disorder of the scurfy mouse. Nat Genet. 2001; 27(1):68-73. (Biology). View Reference
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Lauer FT, Denson JL, Beswick E, Burchiel SW. Intracellular Cytokine Detection by Flow Cytometry in Surface Marker-Defined Human Peripheral Blood Mononuclear T Cells.. Curr Protoc Toxicol. 2017; 73:18.19.1-18.19.14. (Clone-specific: Flow cytometry). View Reference
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Roncador G, Brown PJ, Maestre L, et al. Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level. Eur J Immunol. 2005; 35(6):1681-1691. (Immunogen: Flow cytometry, Immunohistochemistry, Western blot). View Reference
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Wildin RS, Ramsdell F, Peake J, et al. X-linked neonatal diabetes mellitus, enteropathy and endocrinopathy syndrome is the human equivalent of mouse scurfy. Nat Genet. 2001; 27(1):18-20. (Biology). View Reference
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
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