Clone:
REA568
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
IL2RA, IL2R, Ly-43, p55

Extended validation for CD25 Antibody, anti-mouse, 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 clonesOverlap in epitope recognition with REA568
3C7+
7D4++
PC61-
7D4++
Cells were incubated with an excess of purified unconjugated CD25 (REA568) 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 (REA568). 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 (REA568). 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 (REA568). 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, REAfinity™

Overview

Clone REA568 recognizes the mouse CD25 antigen, the α chain of the IL-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 impede the binding of IL-2 to its receptor. In combination with CD4, CD25 antibody can be used to identify regulatory T cells (Tregs).
Additional information: Clone REA568 displays negligible binding to Fc receptors.

Alternative names

IL2RA, IL2R, Ly-43, p55

Detailed product information

Technical specifications

CloneREA568
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD25
Alternative names of antigenIL2RA, IL2R, Ly-43, p55
Molecular mass of antigen [kDa]28
Distribution of antigenT cells, B cells, dendritic cells
Entrez Gene ID16184
RRIDAB_2751464, AB_2752031, AB_2751996, AB_2752187, AB_2752168, AB_2752188, AB_2752169, AB_2811434, AB_2811570, AB_2811565, AB_2751498

Resources for CD25 Antibody, anti-mouse, REAfinity™

Certificates

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

References for CD25 Antibody, anti-mouse, REAfinity™

Publications

  1. Miller, J. et al. (1985) Nucleotide sequence and expression of a mouse interleukin 2 receptor cDNA. J. Immunol. 134(6): 4212-4217
  2. Read, S. et al. (2000)
    Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25
    +
    CD4
    +
    regulatory cells that control intestinal inflammation.
    J. Exp. Med. 192(2): 295-302
  3. Taniguchi, T. et al. (1993) The IL-2/IL-2 receptor system: a current overview. Cell 73(1): 5-8

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