产品详细信息

Description
The eBioRDR5 monoclonal antibody reacts with human CD127 (Interleukin-7 Receptor alpha).

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

Applications Tested
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 (0.8 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.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 μL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 μL of CellBlox Plus Blocking Buffer per 100 μL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Excitation: 493 nm; Emission: 726 nm; Laser: 488 nm (Blue) Laser

靶标信息

CD127, also known as the Interleukin-7 receptor alpha chain (IL-7Ralpha), is a type I glycoprotein with a molecular weight of 75-80 kDa. It forms a multi-functional receptor complex with CD132, the common gamma chain, to create the IL-7 receptor (IL-7R), which is crucial for lymphopoiesis. The active IL-7 receptor is an alpha/gamma chain heterodimer, where the gamma chain also associates with the interleukin-2 receptor and primarily activates signal transduction, while the alpha chain determines specific signaling events through its association with cytoplasmic signaling molecules. CD127 is expressed by immature B cells through the early pre-B stage, thymocytes during various stages of development, and most mature T cells, with transient down-regulation upon activation. It promotes the proliferation of precursor B cells, thymocytes, T cell progenitors, and mature CD4+ and CD8+ T cells. Binding of IL-7 to CD127 results in signal transduction through several tyrosine kinase pathways, including the Jak/STAT pathway, and is indispensable for lymphocyte development and the control of homeostatic proliferation of T cells in the periphery. Additionally, IL-7R signaling is involved in the regulation of T cell receptor (TCR) locus rearrangement in gamma delta T cells. Interestingly, CD127 expression is down-regulated on CD4+CD25+ regulatory T cells (T regs). While CD4 and CD25 co-expression is commonly used to identify T regs, this method may include cells without suppressive activity. CD4+CD25+ cells that have down-regulated CD127 expression are significantly enriched for the regulatory T cell population, as defined by the expression of the T reg-specific transcription factor Foxp3 and their suppressive activity in vitro. Diseases associated with CD127 dysfunction include severe combined immunodeficiency and T cell negative/B cell negative/NK positive severe combined immunodeficiency.