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Blocking of IL-7-induced lymphocyte proliferation. 2E8 cells (mouse immature B lymphocyte cell line) were cultured for 48 hours in the presence of recombinant mouse IL-7 (blue histogram), with mAb SB/14 added 30 minutes prior to IL-7 (red histogram), or in the absence of IL-7 (green histogram). Proliferation was detected by incorporation of bromodeoxyuridine (BrdU) using the BrdU Flow Kit, Cat. No. 559619. Flow cytometry was performed on a FACScan™ (BDIS, San Jose, CA).
BD Pharmingen™ Purified NA/LE 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
Flow cytometry: Since this antigen is expressed at low density on lymphoid cell surfaces, it may be desirable to amplify staining by using a bright second-step reagent. If a three-step method is employed, the use of anti-Mouse CD16/CD32, Mouse Fc Block™ (Cat. No. 553141) is recommended to reduce non-specific staining. Special precaution should be taken to utilize a second-step antibody which does not crossreact with 2.4G2 mAb (rat IgG2b, κ), such as biotinylated anti-rat IgG2a (RG7/1.30, Cat. No. 553894) followed by PE Streptavidin (Cat. No. 554061).
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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The SB/14 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 SB/14 mAb neutralizes IL-7-induced proliferation of the IL-7-dependent immature B-lymphocyte line 2E8 (detected by BrdU incorporation).
Development References (9)
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Akashi K, Kondo M, Weissman IL. Role of interleukin-7 in T-cell development from hematopoietic stem cells. Immunol Rev. 1998; 165:13-28. (Biology). View Reference
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Borghesi LA, Yamashita Y, Kincade PW. Heparan sulfate proteoglycans mediate interleukin-7-dependent B lymphopoiesis. Blood. 1999; 93(1):140-148. (Immunogen). 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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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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Sudo T, Nishikawa S, Ohno N, et al. Expression and function of the interleukin 7 receptor in murine lymphocytes. Proc Natl Acad Sci U S A. 1993; 90(19):9125-9129. (Biology). View Reference
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