CD127 Monoclonal Antibody (eBioRDR5), Super Bright™ 780, eBioscience™
Isotype: IgG1, kappa
Format: Super Bright™ 780
Application: Flow Cytometry
Tested Dilution: 5 µL (0.5 µg)/test
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!
Formulation: PBS, pH 7.2, containing 0.09% sodium azide
Purification: Affinity chromatography
Data Sheet: TDS
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, in vitro.
Binding of the eBioRDR5 monoclonal antibody to PBMCs is blocked by pre-incubation of the cells with recombinant human IL-7 (cat. 14-8079).
Applications Reported: This eBioRDR5 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This eBioRDR5 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may 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 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 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 # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.
In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).
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 # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) 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: 405 nm; Emission: 780 nm; Laser: Violet Laser
Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.