CD127 Monoclonal Antibody (A7R34), PE-eFluor™ 610, eBioscience™, Invitrogen™

Description

Description: The A7R34 monoclonal antibody reacts with mouse CD127, the high affinity alpha subunit of the mouse IL-7 receptor. IL-7 receptor alpha chain is expressed by immature B cells in the bone marrow, double-negative (CD4-CD8-), single-positive (CD4+ and CD8+), but not double-positive (CD4+CD8+) thymocytes. In the periphery, mature T cells express CD127 at low level. A7R34 inhibits binding of IL-7 to its receptor and has been used in in vivo and in vitro studies to elucidate the role of IL-7 in T and B cell development and activation. Binding of A7R34 blocks the binding of SB/199, another antibody which recognizes mouse CD127. Applications Reported: This A7R34 antibody has been reported for use in flow cytometric analysis. Applications Tested: This A7R34 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.25 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PE-eFluor 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red).  Please make sure that your instrument is capable of detecting this fluorochrome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product No. 00-8222-49) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser 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.