Clone:
REA945
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
IL2RA, IDDM10, IL2R, TCGFR, p55, Tac, IL-2Rα

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 clonesOverlap in epitope recognition with REA945
4E3++
3G10-
REA570-
2A3-
BC96-
M-A251++
Cells were incubated with an excess of purified unconjugated CD25 (REA945) 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. 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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD25 (REA945). 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 (REA945). 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 (REA945). 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 REA945 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. CD25 plays an essential role in the regulation of immune tolerance by controlling regulatory T cells activity. The CD25 antigen contains three epitope regions called A, B, and C. Clone REA945 recognizes epitope region B.
Additional information: Clone REA945 displays negligible binding to Fc receptors.

Alternative names

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

Detailed product information

Technical specifications

CloneREA945
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, non-human primate
Cross-reactivity
cynomolgus monkey (
Macaca fascicularis
)
,
rhesus monkey (
Macaca mulatta
)
AntigenCD25
Alternative names of antigenIL2RA, IDDM10, IL2R, TCGFR, p55, Tac, IL-2Rα
Molecular mass of antigen [kDa]28
Distribution of antigenB cells, granulocytes, lymphocytes, macrophages, monocytes, NK cells, osteoclasts, T cells, basophils, thymocytes
Entrez Gene ID3559
RRIDAB_2727130, AB_2727075, AB_2727076, AB_2727131, AB_2727077, AB_2727464, AB_2727402, AB_2801690, AB_2801689, AB_2727129, AB_2727074

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

  1. 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
  2. Shevach, E. M. (2001)
    Certified professionals: CD4
    +
    CD25
    +
    suppressor T cells.
    J. Exp. Med. 193(11): F41-F46
  3. Maloy, K. J. et al. (2001) Regulatory T cells in the control of immune pathology. Nat. Immunol. 2(9): 816-822
  4. 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
  5. 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

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