CD127 Antibody, anti-human, REAfinity™

Name: CD127 Antibody, anti-human, REAfinity™
Availability: InStock

CD127 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA614

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MC

Alternative names:

IL-7R, IL-7Rα, IL-17Ralpha, CDW127, ILRA

Flow cytometry applications forCD127 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
Vio Bright B515
Vio Bright FITC
Vio Bright V423

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-113-975, 130-113-413

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD127 antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD3 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

Mass cytometry applications forCD127 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-122-294

Extended validation for CD127 Antibody, anti-human, REAfinity™

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with REA614
HIL-7R-M21	++
eBioRDR5	-
A019D5	++
MB15-18C9	++
Cells were incubated with an excess of purified unconjugated CD127 (REA614) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for CD127. Human peripheral blood mononuclear cells (PBMCs) were stained with CD127 antibodies and with a suitable counterstaining. As a control, CD127 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD127. Human peripheral blood mononuclear cells (PBMCs) were stained with CD127 antibodies and with a suitable counterstaining. As a control, CD127 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD127. Human peripheral blood mononuclear cells (PBMCs) were stained with CD127 antibodies and with a suitable counterstaining. As a control, CD127 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD127. Human peripheral blood mononuclear cells (PBMCs) were stained with CD127 antibodies and with a suitable counterstaining. As a control, CD127 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD127. Human peripheral blood mononuclear cells (PBMCs) were stained with CD127 antibodies and with a suitable counterstaining. As a control, CD127 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD127. Human peripheral blood mononuclear cells (PBMCs) were stained with CD127 antibodies and with a suitable counterstaining. As a control, CD127 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Fixation data

To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using CD127 (REA614). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD127 (REA614). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD127 Antibody, anti-human, REAfinity™

Overview

Clone REA614 recognizes the CD127 antigen, which is the α-chain of the interleukin 7 (IL-7) receptor, a type I membrane glycoprotein. Signaling of IL-7 through the IL-7R requires both IL-7Rα and the common cytokine gamma chain (γc). CD127 can be identified on immature B cells through the early pre–B stage, on thymocytes, and on most mature T cells with transient down-regulation upon activation. On regulatory T cells CD127 is absent and its expression is inversely correlated with FoxP3 expression and suppressive function. CD127 is also used by thymic stromal derived lymphopoietin (TSLP) as part of a complex.

Additional information: Clone REA614 displays negligible binding to Fc receptors.

Alternative names

IL-7R, IL-7Rα, IL-17Ralpha, CDW127, ILRA

Detailed product information

Technical specifications

Clone	REA614
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD127
Alternative names of antigen	IL-7R, IL-7Rα, IL-17Ralpha, CDW127, ILRA
Molecular mass of antigen [kDa]	50
Distribution of antigen	T cells
Entrez Gene ID	3575
RRID	AB_2726163, AB_2733758, AB_2733759, AB_2726438, AB_2726161, AB_2733197, AB_2733198, AB_2726439, AB_2726162, AB_2783927, AB_2783926, AB_2811440, AB_2811433, AB_2921782, AB_2921784, AB_2801875, AB_2921932, AB_2921907, AB_2726440

Resources for CD127 Antibody, anti-human, REAfinity™

Documents and Protocols

Brochures/posters

REAfinity™ Antibodies for mass cytometry - Recombinantly engineered to increase reproducibility

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD127 Antibody, anti-human, REAfinity™

Publications

Fry, T. J. and Mackall, C. L. (2002) Interleukin-7: from bench to clinic. Blood 99: 3892-3904
Seddiki, N. et al. (2006) Expression of interleukin (IL)-2 and IL-7 receptors discriminates between human regulatory and activated T cells. J. Exp. Med. 203: 1693-1700
Liu, W. et al. (2006)
CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4
⁺
T reg cells.
J. Exp. Med. 203: 1701-1711
Sudo, T. et al. (1993) Expression and function of the interleukin 7 receptor in murine lymphocytes. Proc. Natl. Acad. Sci. U.S.A. 90: 9125-9129
Cupedo, T. et al. (2005) Development and activation of regulatory T cells in the human fetus. Eur. J. Immunol. 35: 383-390
Armitage, R. J. et al. (1991) Expression of receptors for interleukin 4 and interleukin 7 on human T cells. Adv. Exp. Med. Biol. 292: 121-130

Applications

Extended validation