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Two-color flow cytometric analysis of CD127 expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826) and either BD Horizon™ BUV737 Rat IgG2b, κ Isotype Control (Cat. No. 612762; Left Plot) or BD Horizon BUV737 Rat Anti-Mouse CD127 antibody (Cat. No. 612841; Right Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. Two-color flow cytometric contour plots showing the correlated expression patterns of CD127 (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BUV737 Rat Anti-Mouse 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
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.
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) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Ultraviolet 737 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 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 http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The SB/199 monoclonal antibody specifically binds to mouse CD127 which is also known as, the Interleukin-7 Receptor alpha chain (IL-7Rα). CD127 associates with CD132 (common γ chain) to form a high-affinity signaling IL-7R complex that mediates the biological effects of IL-7. CD127 can also complex with the Thymic Stromal Lymphopoietin Receptor (TSLPR) subunit to bind and mediate cellular responses to TSLP. CD127 is a 65-75 kDa type-I transmembrane protein that belongs to the hematopoietin receptor and the Ig gene superfamilies. Mice lacking CD127 display profoundly impaired development of the B and T lymphoid cell lineages, but display no obvious nonlymphoid abnormalities. IL-7Rα is expressed on common lymphoid progenitors and early stages of B lineage development in the bone marrow, on the earliest thymocyte progenitors, on CD4-CD8- double-negative and CD4+ and CD8+ single-positive thymocytes, and on most peripheral T lymphocytes. Intestinal intraepithelial lymphocytes with low-density γδ TCR expression upregulate CD127 expression in response to IL-2, which may be secreted by neighboring αβ TCR+ T cells.
The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter. Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.
Development References (10)
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Brugnera E, Bhandoola A, Cibotti R, et al. Coreceptor reversal in the thymus: signaled CD4+8+ thymocytes initially terminate CD8 transcription even when differentiating into CD8+ T cells. Immunity. 2000; 13(1):59-71. (Biology). View Reference
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Faust EA, Saffran DC, Toksoz D, Williams DA, Witte ON. Distinctive growth requirements and gene expression patterns distinguish progenitor B cells from pre-B cells. J Exp Med. 1993; 177(4):915-923. (Biology). View Reference
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Fujihashi K, Kawabata S, Hiroi T, et al. Interleukin 2 (IL-2) and interleukin 7 (IL-7) reciprocally induce IL-7 and IL-2 receptors on gamma delta T-cell receptor-positive intraepithelial lymphocytes. Proc Natl Acad Sci U S A. 1996; 93(8):3613-3618. (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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Henderson AJ, Narayanan R, Collins L, Dorshkind K. Status of kappa L chain gene rearrangements and c-kit and IL-7 receptor expression in stromal cell-dependent pre-B cells. J Immunol. 1992; 149(6):1973-1979. (Biology). View Reference
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Igarashi H, Gregory SC, Yokota T, Sakaguchi N, Kincade PW. Transcription from the RAG1 locus marks the earliest lymphocyte progenitors in bone marrow.. Immunity. 2002; 17(2):117-30. (Clone-specific: Flow cytometry). View Reference
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Noguchi M, Nakamura Y, Russell SM, et al. Interleukin-2 receptor gamma chain: a functional component of the interleukin-7 receptor. Science. 1993; 262(5141):1877-1880. (Biology). View Reference
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Peschon JJ, Morrissey PJ, Grabstein KH, et al. Early lymphocyte expansion is severely impaired in interleukin 7 receptor-deficient mice. J Exp Med. 1994; 180(5):1955-1960. (Biology). View Reference
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Yamashita Y, Oritani K, Miyoshi EK, Wall R, Bernfield M, Kincade PW. Syndecan-4 is expressed by B lineage lymphocytes and can transmit a signal for formation of dendritic processes. J Immunol. 1999; 162(10):5940-5948. (Clone-specific: Flow cytometry). View Reference
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