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CD127 Monoclonal Antibody (A7R34), NovaFluor™ Red 700, eBioscience™
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FIGURE: 1 / 3
CD127 Antibody (M007T02R03-A) in Flow
C57BL/6 mouse splenocytes were unstained (left) or stained with 1.0 µg of CD127 Monoclonal Antibody, NovaFluor Red 700 (right). All cells were co-stained with CD3e Monoclonal Antibody, Super Bright 436 (Product # 2-0031-82). Total viable cells were used for analysis, as determined by LIVE/DEAD Fixable Blue Viability Dye (Product # L34963). Data was acquired in the R3 channel on a 5-laser Cytek Aurora for the NovaFluor Red 700 conjugate.
Product Details
M007T02R03-A
Product Specifications
Species Reactivity
Mouse
Host/Isotype
Rat
/ IgG2a, kappa
Class
Monoclonal
Type
Antibody
Clone
A7R34
Conjugate
Excitation/Emission Max
637/701 nm
View spectra
Form
Liquid
Concentration
0.1 mg/mL
Amount
25 µg
Purification
Affinity chromatography
Storage buffer
PBS, pH 7.2
Contains
0.09% sodium azide
Storage conditions
4°C, store in dark, DO NOT FREEZE!
RRID
Product Specific Information
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.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
Applications Tested
The A7R34 antibody has been tested by flow cytometric analysis of mouse thymocytes and splenocytes. This can be used at less than or equal to 1.0 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.
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 (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, 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: 637 nm; Emission: 701 nm; Laser: 633-640 nm (Red) Laser
Target Information
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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Bioinformatics
Protein Aliases: CD127; IL 7 receptor; IL 7R a; IL 7R subunit alpha; IL 7R α; IL 7Ra; IL 7Rα; IL-7 receptor alpha chain; IL-7 receptor subunit alpha; IL-7R; IL-7R subunit alpha; IL-7R-alpha; IL-7RA; IL7 receptor; IL7 receptor subunit alpha; IL7R a; IL7R alpha; IL7R subunit alpha; IL7R α; IL7Rα; interleukin 7 receptor alpha chain; Interleukin-7 receptor subunit alpha; Interleukin7 receptor subunit alpha; MGC107557; sCD127; soluble IL 7 receptor; soluble IL 7R
Gene Aliases: CD127; IL-7Ralpha; Il7r
UniProt ID: (Mouse) P16872
Entrez Gene ID: (Mouse) 16197
cell morphogenesis
negative regulation of T cell mediated cytotoxicity
immunoglobulin production
regulation of cell size
positive regulation of gene expression
cell growth
T cell differentiation
positive regulation of T cell differentiation in thymus
B cell proliferation
lymph node development
homeostasis of number of cells
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