CD25 Antibody, anti-mouse

Name: CD25 Antibody, anti-mouse
Availability: InStock

CD25 Antibody, anti-mouseExtended Validation

Clone:

7D4

Type of antibody:

Primary antibodies

Isotype:

rat IgMκ

Applications:

FC, MICS, IF, IHC

Alternative names:

IL2RA, IL2R, Ly-43, IL-2Rα, p55, Tac

Flow cytometry applications forCD25 Antibody, anti-mouse

APC
Biotin
PE
PE-Vio 770
Vio Bright FITC

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:

130-116-496, 130-116-532

Protocols:

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

Description:

Mouse splenocytes were stained with CD25 antibodies as well as with CD4 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

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

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-116-496, 130-116-532

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD25 Antibody, anti-mouse

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 7D4
3C7	+
REA568	++
PC61	-
Cells were incubated with an excess of purified unconjugated CD25 (7D4) 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. Splenocytes from BALB/c mice were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25antibody 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. Splenocytes from BALB/c mice were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25antibody 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. Splenocytes from BALB/c mice were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25antibody 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. Splenocytes from BALB/c mice were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25antibody 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. Splenocytes from BALB/c mice were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25antibody 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. Splenocytes from BALB/c mice were stained with CD25 antibodies and with a suitable counterstaining. As a control, CD25antibody 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 (7D4). 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 (7D4). 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-mouse

Overview

The CD25 antibody recognizes the α chain of the interleukin 2 receptor (IL-2R). IL-2Rα associates with the β chain (CD122) and the γ chain (CD132) to form the functional IL-2R complex. IL-2Rα is expressed on CD4

⁺

CD25

⁺

regulatory T cells, activated T and B cells, and to a lesser extent on activated dendritic cells. It is also transiently expressed during T and B cell development. Binding of the CD25 antibody does not inhibit the binding of IL-2 to its receptor. In combination with CD4, CD25 antibody can be used to identify regulatory T cells (Tregs).

Alternative names

IL2RA, IL2R, Ly-43, IL-2Rα, p55, Tac

Detailed product information

Technical specifications

Clone	7D4
Clonality	monoclonal
Isotype	rat IgMκ
Isotype control	Isotype Control Antibody, rat IgM
Host	rat
Type of antibody	Primary antibodies
Species	mouse
Antigen	CD25
Alternative names of antigen	IL2RA, IL2R, Ly-43, IL-2Rα, p55, Tac
Molecular mass of antigen [kDa]	28
Distribution of antigen	B cells, macrophages, monocytes, NK cells, osteoblasts, T cells, lymphocytes, basophils, thymocytes
Entrez Gene ID	16184
RRID	AB_2784088, AB_2727571, AB_2727595, AB_2751787, AB_2784091, AB_2784090, AB_2857607, AB_2784089

Resources for CD25 Antibody, anti-mouse

Certificates

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

Applications

Extended validation