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

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 clonesOverlap 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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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
View details
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.
View details
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

Clone7D4
Clonalitymonoclonal
Isotyperat IgMκ
Isotype controlIsotype Control Antibody, rat IgM
Hostrat
Type of antibodyPrimary antibodies
Speciesmouse
AntigenCD25
Alternative names of antigenIL2RA, IL2R, Ly-43, IL-2Rα, p55, Tac
Molecular mass of antigen [kDa]28
Distribution of antigenB cells, macrophages, monocytes, NK cells, osteoblasts, T cells, lymphocytes, basophils, thymocytes
Entrez Gene ID16184
RRIDAB_2784088, AB_2727571, AB_2727595, AB_2751787, AB_2784091, AB_2784090, AB_2857607, AB_2784089

Resources for CD25 Antibody, anti-mouse

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

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