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Multicolor flow cytometric analysis of CD127 expression on human peripheral blood CD4+ T lymphocytes. Human whole blood was stained with APC Mouse Anti-Human CD4 (Cat. No. 565994) and PE Mouse Anti-Human CD25 (Cat. No. 555432/560989) antibodies, and either BD Horizon™ BV480 Mouse IgG1, κ Isotype Control (Cat. No. 565652; Left Plot) or BD Horizon BV480 Mouse Anti-Human CD127 (IL-7 Receptor α) antibody (Cat. No. 566101/566158; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric dot plots showing the correlated expression of CD25 (or Ig Isotype control staining) versus CD127 were derived from CD4+ gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Horizon™ BV480 Mouse Anti-Human CD127
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The hIL-7R-M21 monoclonal antibody specifically binds to the 60-90 kDa glycoprotein, CD127. CD127 is also known as the IL-7 receptor alpha (IL-7Rα) subunit. The IL-7 receptor complex is a heterodimer composed of CD127 and the common gamma chain (γc, CD132), shared by other cytokine receptors (IL-2R, IL-4R, IL-9R, IL-15R, and IL-21R). CD127 is expressed on thymocytes, T- and B-cell progenitors, mature T cells, and some lymphoid and myeloid cells. In vitro experiments show the expression of CD127 is down-regulated following T cell activation. Studies indicate that the IL-7 Receptor plays an important role in the proliferation and differentiation of mature T cells. Recently, it has been shown that low surface expression of CD127, in combination with intermediate to high surface expression of CD25, the α chain of the IL-2 receptor complex, can distinguish between human regulatory and conventional CD4+ T cells in human adult and cord blood, lymph nodes and thymus.
The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set. BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.
Development References (7)
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Appasamy PM. Biological and clinical implications of interleukin-7 and lymphopoiesis. Cytokines Cell Mol Ther. 1999; 5(1):25-39. (Biology: Flow cytometry). View Reference
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Armitage RJ, Ziegler SF, Friend DJ, Park LS, Fanslow WC. Identification of a novel low-affinity receptor for human interleukin-7. Blood. 1992; 79(7):1738-1745. (Immunogen: Flow cytometry). View Reference
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Benjamin D, Sharma V, Knobloch TJ, Armitage RJ, Dayton MA, Goodwin RG. B cell IL-7. Human B cell lines constitutively secrete IL-7 and express IL-7 receptors. J Immunol. 1994; 152(10):4749-4757. (Clone-specific: Flow cytometry). View Reference
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Goodwin RG, Friend D, Ziegler SF et al. Cloning of the human and murine interleukin-7 receptors: demonstration of a soluble form and homology to a new receptor superfamily. Cell. 1990; 60(6):941-951. (Biology). View Reference
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Hofmeister R, Khaled AR, Benbernou N, Rajnavolgyi E, Muegge K, Durum SK. Interleukin-7: physiological roles and mechanisms of action. Cytokine Growth Factor Rev. 1999; 10(1):41-60. (Biology). View Reference
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Liu W, Putnam AL, Xu-Yu Z, et al. CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells. J Exp Med. 2006; 203(7):1701-1711. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Seddiki N, Santner-Nanan B, Martinson J et al. Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells. J Exp Med. 2006; 203(7):1693-1700. (Biology). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.