Clone:
REA918
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, ICC, MC
Alternative names:
IL3RA, IL3R, IL3RAY, IL3RX, IL3RY, hIL-3Ra, IL-3Rα

Extended validation for CD123 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 REA918
AC145++
6H6++
7G3++
32703-
Cells were incubated with an excess of purified unconjugated CD123 (REA918) 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 CD123. Human peripheral blood mononuclear cells (PBMCs) were stained with CD123 antibodies and with a suitable counterstaining. As a control, CD123 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 CD123. Human peripheral blood mononuclear cells (PBMCs) were stained with CD123 antibodies and with a suitable counterstaining. As a control, CD123 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 CD123. Human peripheral blood mononuclear cells (PBMCs) were stained with CD123 antibodies and with a suitable counterstaining. As a control, CD123 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 CD123. Human peripheral blood mononuclear cells (PBMCs) were stained with CD123 antibodies and with a suitable counterstaining. As a control, CD123 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 CD123. Human peripheral blood mononuclear cells (PBMCs) were stained with CD123 antibodies and with a suitable counterstaining. As a control, CD123 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 CD123. Human peripheral blood mononuclear cells (PBMCs) were stained with CD123 antibodies and with a suitable counterstaining. As a control, CD123 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 CD123 (REA918). 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 CD123 (REA918). 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 CD123 (REA918). 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 CD123 Antibody, anti-human, REAfinity™

Overview

Clone REA918 recognizes the human CD123 antigen, a type I transmembrane glycoprotein, which is also known as interleukin 3 (IL-3) receptor α-chain and is the primary low-affinity subunit of the IL-3 receptor. CD123 associates with CD131, the common β-chain of the IL-3, IL-5, and GM-CSF receptor, to form the high-affinity IL-3 receptor. The IL-3 receptor is involved in cell signaling for cell growth and differentiation. In peripheral blood, the CD123 antigen is expressed at high levels only on plasmacytoid dendritic cells and basophilic granulocytes but at low levels also on monocytes, eosinophilic granulocytes, myeloid dendritic cells, and subsets of hematopoietic progenitor cells.
Additional information: Clone REA918 displays negligible binding to Fc receptors.

Alternative names

IL3RA, IL3R, IL3RAY, IL3RX, IL3RY, hIL-3Ra, IL-3Rα

Detailed product information

Technical specifications

CloneREA918
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
rhesus monkey (
Macaca mulatta
)
AntigenCD123
Alternative names of antigenIL3RA, IL3R, IL3RAY, IL3RX, IL3RY, hIL-3Ra, IL-3Rα
Molecular mass of antigen [kDa]41
Distribution of antigendendritic cells, granulocytes, mast cells, megakaryocytes, monocytes, basophils, eosinophils, hematopoietic stem and progenitor cells, myeloid dendritic cells, plasmacytoid dendritic cells
Entrez Gene ID3563
RRIDAB_2726967, AB_2727025, AB_2726968, AB_2727026, AB_2726969, AB_2727030, AB_2726973, AB_2727031, AB_2726974, AB_2727032, AB_2726975, AB_2727027, AB_2726970, AB_2727028, AB_2726971, AB_2727029, AB_2726972, AB_2727023, AB_2726966, AB_2751497, AB_2751463, AB_2801878, AB_2727024

Resources for CD123 Antibody, anti-human, REAfinity™

References for CD123 Antibody, anti-human, REAfinity™

Publications

  1. Miyajima, A. et al. (1993) Receptors for granulocyte-macrophage colony-stimulating factor, interleukin-3, and interleukin-5. Blood 82(7): 1960-1974
  2. Chen, J. et al. (2009) A new isoform of interleukin-3 receptor {alpha} with novel differentiation activity and high affinity binding mode. J. Biol. Chem. 284(9): 5763-5773
  3. Choi, K.-D. et al. (2011) Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells. Nat. Protoc. 6(3): 296-313

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