CD25 Antibody, anti-human

Name: CD25 Antibody, anti-human
Availability: InStock

CD25 Antibody, anti-humanExtended Validation

Clone:

4E3

Type of antibody:

Primary antibodies

Isotype:

mouse IgG2bκ

Applications:

FC, MICS, IF, IHC

Alternative names:

IL2RA, IDDM10, IL2R, TCGFR, p55, Tac, IL-2Rα

Flow cytometry applications forCD25 Antibody, anti-human

APC
Biotin
PE
Vio Bright FITC

Conjugate:

APC

Recommended dilution:

1:50 (1:25 for CD25 MicroBead-labeled cells)

Tested:

QC tested

Products used:

130-113-842, 130-113-280

Protocols:

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

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD25 antibodies or with the corresponding isotype control antibodies (left images) as well as with CD4 antibodies and analyzed by flow cytometry 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.

MICS (MACSima Imaging Cyclic Staining) applications forCD25 Antibody, anti-human

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-113-842, 130-113-280

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD25 Antibody, anti-human

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 (4E3). 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 (4E3). 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

Overview

The CD25 antibody recognizes the human CD25 antigen, a 28 kDa glycoprotein, also known as the low-affinity interleukin-2 receptor alpha chain (IL-2Rα). CD25 is expressed on activated T and B cells, on macrophages, and on a subset of non-activated CD4

⁺

regulatory T cells.

The CD25 antigen contains three epitope regions called A, B, and C. This CD25 antibody recognizes epitope region B.

Alternative names

IL2RA, IDDM10, IL2R, TCGFR, p55, Tac, IL-2Rα

Detailed product information

Technical specifications

Clone	4E3
Clonality	monoclonal
Isotype	mouse IgG2bκ
Isotype control	Isotype Control Antibody, mouse IgG2b
Host	mouse
Type of antibody	Primary antibodies
Species	human, non-human primate
Cross-reactivity	rhesus monkey ( Macaca mulatta ) , cynomolgus monkey ( Macaca fascicularis )
Antigen	CD25
Alternative names of antigen	IL2RA, IDDM10, IL2R, TCGFR, p55, Tac, IL-2Rα
Molecular mass of antigen [kDa]	28
Distribution of antigen	T cells, B cells, NK cells, lymphocytes, macrophages, monocytes, osteoblasts, thymocytes, basophils
Entrez Gene ID	3559
RRID	AB_2734062, AB_2733789, AB_2733790, AB_2726345, AB_2726068, AB_2733510, AB_2733511, AB_2734061

Resources for CD25 Antibody, anti-human

Certificates

Please follow this

link

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

References for CD25 Antibody, anti-human

Publications

Sakaguchi, S. et al. (1995) Immunologic self-tolerance maintained by activated T cells expressing IL-2 receptor α-chains (CD25). Breakdown of a single mechanism of self-tolerance causes various autoimmune diseases. J Immunol 155: 1151-1164
Lamprecht, B. et al. (2008) Aberrant expression of the Tʜ2 cytokine IL-21 in Hodgkin lymphoma cells regulates STAT3 signaling and attracts Treg cells via regulation of MIP-3alpha. Blood 112(8): 3339-3347
Maloy, K. J. et al. (2001) Regulatory T cells in the control of immune pathology. Nat Immunol 2(9): 816-822
Shevach, E. M. (2001)
Certified professionals: CD4
⁺
CD25
⁺
suppressor T cells.
J. Exp. Med. 193(11): F41-F46
Chaput, N. et al. (2013)
Phase I clinical trial combining imatinib mesylate and IL-2: HLA-DR
⁺
NK cell levels correlate with disease outcome.
Oncoimmunology 2(2): e23080

Applications

Extended validation