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Flow cytometric analysis of CD127 expression on human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ BV650 Mouse Anti-Human CD127 antibody (Cat. No. 563225; solid line histogram) or with a BD Horizon™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV650 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
Caution: Investigators should note that usage with PBMC, despite titration, may result in higher background staining and/or uncharacteristic flow cytometric profiles. In such cases, investigators are highly encouraged to consider using other formats, such as PE or FITC. Whole blood may also be used as an alternative.
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. 563794/566349).
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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
- BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- 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™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon™ BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon™ BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (5)
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Akashi K, Traver D, Kondo M, Weissman IL. Lymphoid development from hematopoietic stem cells. Int J Hematol. 1999; 69(4):217-226. (Biology). View Reference
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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). 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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In: Fitzgerald KA, Callard RE. The cytokine factsbook., 2nd ed. / Katherine A. Fitzgerald ... [et al.].. San Diego: Academic Press; 2001:75.
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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.