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Two-color flow cytometric analysis of FoxP3 expression in resting Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stained with BD Horizon™ BUV395 Mouse Anti-Human CD4 antibody (Cat. No. 563550). The cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574) followed by staining with either BD Horizon™ RB670 Mouse IgG1, κ Isotype Control (Cat. No. 571784; Left Plot) or BD Horizon™ RB670 Mouse Anti-Human FoxP3 antibody (Cat. No. 571930/571932; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of FoxP3 versus CD4 [or Ig Isotype control staining] was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Samples were acquired using the BD FACSymphony™ A5 SE Cell Analyzer System and were spectrally unmixed with autofluorescence using FlowJo™ v10.10 Software.
BD Horizon™ RB670 Mouse Anti-Human FoxP3
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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Companion Products
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.
Development References (7)
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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: Intracellular Staining/Flow Cytometry). View Reference
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Law JP, Hirschkorn DF, Owen RE, Biswas HH, Norris PJ, Lanteri MC. The importance of Foxp3 antibody and fixation/permeabilization buffer combinations in identifying CD4+CD25+Foxp3+ regulatory T cells.. Cytometry A. 2009; 75(12):1040-50. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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Mailer RKW. Alternative Splicing of FOXP3-Virtue and Vice.. Front Immunol. 2018; 9:530. (Biology). 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: Immunohistochemistry, Intracellular Staining/Flow Cytometry, Western blot). View Reference
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Ruitenberg JJ, Boyce C, Hingorani R, Putnam A, Ghanekar SA. Rapid assessment of in vitro expanded human regulatory T cell function.. J Immunol Methods. 2011; 372(1-2):95-106. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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Sadlon TJ, Wilkinson BG, Pederson S, et al. Genome-wide identification of human FOXP3 target genes in natural regulatory T cells. J Immunol. 2010; 185(2):1071-1081. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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Santegoets SJ, Dijkgraaf EM, Battaglia A, et al. Monitoring regulatory T cells in clinical samples: consensus on an essential marker set and gating strategy for regulatory T cell analysis by flow cytometry.. Cancer Immunol Immunother. 2015; 64(10):1271-86. (Clone-specific: Intracellular Staining/Flow Cytometry). View Reference
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