CD25 Antibody, anti-human, REAfinity™

Name: CD25 Antibody, anti-human, REAfinity™
Availability: InStock

CD25 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA570

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MC, 3D-IF, IF, IHC

Alternative names:

IL2RA, IDDM10, IL2R, TCGFR, p55, Tac

Flow cytometry applications forCD25 Antibody, anti-human, REAfinity™

APC
APC-Vio 770
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright B515
Vio Bright R667

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-113-846, 130-113-284

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs), overnight stimulated with CytoStim, were stained with CD25 antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD4 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

3D-Immunofluorescence applications forCD25 Antibody, anti-human, REAfinity™

Vio R667

Conjugate:

Vio R667

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Products used:

130-131-142

Tissue clearing:

MACS^® Clearing Kit

Compatible applications:

IF, IHC

Protocols:

Immunostaining and clearing of primary human tumors and patient-derived xenografts for 3D imaging ...

Description:

3D-immunofluorescence analysis of human colorectal cancer fixed with 4% PFA and subjected to whole-mount immunostaining with CD25 Antibody, anti human, Vio R667 (red). The sample was processed and optically cleared using the MACS^® Clearing Kit and imaged with an UltraMicroscope Blaze light sheet instrument. Sample detail image (A), overview image (B) and 3D-rendered video (C). Scale bars: 100 µm (A), 500 µm (B, C).

Mass cytometry applications forCD25 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Reported:

32676612,

Products used:

130-122-302

Extended validation for CD25 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 REA570
2A3	++
M-A251	-
4E3	-
BC96	++
M-A251	-
3G10	++
REA945	-
Cells were incubated with an excess of purified unconjugated CD25 (REA570) 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25. Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25 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 CD25 (REA570). 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 CD25 (REA570). 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 CD25 Antibody, anti-human, REAfinity™

Overview

Clone REA570 recognizes the human CD25 glycoprotein, also known as the low-affinity interleukin-2 receptor alpha chain (IL-2Rα). It associates with IL-2 receptor β (CD122) and γ chains (CD132) to form a high affinity IL-2R complex. CD25 is expressed on activated T and B cells, on macrophages, and on a subset of non-activated CD4

⁺

regulatory T cells. Clone REA570 recognizes epitope region A.

Additional information: Clone REA570 displays negligible binding to Fc receptors

Alternative names

IL2RA, IDDM10, IL2R, TCGFR, p55, Tac

Detailed product information

Technical specifications

Clone	REA570
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
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , olive baboon ( Papio anubis )
Antigen	CD25
Alternative names of antigen	IL2RA, IDDM10, IL2R, TCGFR, p55, Tac
Molecular mass of antigen [kDa]	55
Distribution of antigen	T cells, B cells, macrophages
Entrez Gene ID	3559
RRID	AB_2733792, AB_2726346, AB_2726069, AB_2726777, AB_2726693, AB_2784093, AB_2784092, AB_2726347, AB_2726070, AB_2819448, AB_2819444, AB_2819496, AB_2819488, AB_2801883, AB_2904878, AB_2904877, AB_2733791

Resources for CD25 Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

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

References for CD25 Antibody, anti-human, REAfinity™

Publications

Lee, S. H. et al. (2012) Cutting edge: a novel mechanism bridging innate and adaptive immunity: IL-12 induction of CD25 to form high-affinity IL-2 receptors on NK cells. J Immunol 189(6): 2712-2716
Piccirillo, C. A. et al. (2006)
Development and function of naturally occurring CD4
⁺
CD25
⁺
regulatory T cells.
J. Leukoc. Biol. 3: 458-470
Goudy, K. et al. (2013) Human IL2RA null mutation mediates immunodeficiency with lymphoproliferation and autoimmunity. Clin. Immunol. 146(3): 248-261

Applications

Extended validation