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Flow cytometric analysis of CD127 expression on human peripheral blood leucocytes. Human whole blood was stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot) or BD Horizon R718 Mouse Anti-Human CD127 antibody (Cat. No. 566967/566968; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A two-parameter pseudocolor density plot showing the correlated expression of CD127 (or Ig Isotype control staining) versus side light-scatter (SSC) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human CD127
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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
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.
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.
- 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.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
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 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.
Development References (8)
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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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Sasidharan Nair V, Toor SM, Taouk G, et al. Pembrolizumab Interferes with the Differentiation of Human FOXP3(+)-Induced T Regulatory Cells, but Not with FOXP3 Stability, through Activation of mTOR. J Immunol. 2020; 204(1):199-211. (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.