CD127 Monoclonal Antibody (eBioRDR5), Super Bright™ 600, eBioscience™, Invitrogen™

Description

The eBioRDR5 monoclonal antibody reacts with human CD127 (Interleukin-7 Receptor alpha). CD127 complexes with CD132, also known as the common gamma chain (gamma c), to form the multi-functional IL-7 receptor (IL-7R). CD127 is a type I glycoprotein with a molecular weight of 75-80 kDa and is expressed by immature B cells through the early pre-B stage, by thymocytes during several stages of their development, and on most mature T cells, with transient down-regulation upon activation. Binding of IL-7 results in signal transduction which occurs through several tyrosine kinase pathways including the Jak/STAT pathway. IL-7 is indispensible for the development of lymphocytes, and the control of homeostatic proliferation of T-cells in the periphery. In addition, IL-7R signaling is know to be involved in the regulation of T cell receptor (TCR) locus rearrangement in gamma delta T cells. Interestingly, recently it has been demonstrated that CD127 expression is down-regulated on CD4+CD25+ regulatory T cells (T regs). While the co-expression of CD4 and CD25 has become widely used as an indicator of T regs, this method of identification may also include cells without suppressive activity.

It has clearly been shown that CD4+CD25+ cells that have down-regulated the expression of CD127 are significantly more highly-enriched for the regulatory T population, as defined by expression of the T reg-specific transcription factor Foxp3 and the suppressive activity of these cells, <i>in vitro</i>. Binding of the eBioRDR5 monoclonal antibody to PBMCs is blocked by pre-incubation of the cells with recombit human IL-7 (cat. 14-8079). The antibody eBioRDR5 has been published to crossreact to rhesus CD127. This eBioRDR5 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 μL (0.5 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final volume of 100 μL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Super Bright 600 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 600 nm. We recommend using a 610/20 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product No. SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for upto 3 days in the dark at 2-8°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorochrome performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 600 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company. CD127 (Interleukin-7, IL-7) is a glycoprotein involved in the regulation of lymphopoiesis. The response of cells to CD127 is dependent on the presence of the interleukin 7 receptor (IL7R); the active receptor is an alpha/gamma chain heterodimer. CD127 consists of an alpha chain and a gamma chain. The gamma(c) chain, which also associates with the interleukin-2 receptor, serves primarily to activate signal transduction by the IL7R complex, while the alpha chain of IL7R determines specific signaling events through its association with cytoplasmic signaling molecules. CD127 promotes the proliferation of precursor B cells, thymocytes, T cell progenitors, and mature CD4+ and CD8+ T cells. The biological effects of IL7 are mediated by the binding of IL7 to the specific cell surface receptor. Diseases associated with CD127 dysfunction include severe combined immunodeficiency and T cell negative/B cell negative/NK positive severe combined immunodeficiency.