Clone:
REA343
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
CDw125, Il5r, IL-5RA

Extended validation for CD125 (IL-5Rα) 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 REA343
T21++
Cells were incubated with an excess of purified unconjugated CD125 (IL-5Rα) (REA343) 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 CD125 (IL-5Rα). Splenocytes from BALB/c mice were stained with CD125 (IL-5Rα) antibodies and with a suitable counterstaining. As a control, CD125 (IL-5Rα) 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 CD125 (IL-5Rα). Splenocytes from BALB/c mice were stained with CD125 (IL-5Rα) antibodies and with a suitable counterstaining. As a control, CD125 (IL-5Rα) 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 CD125 (IL-5Rα). Splenocytes from BALB/c mice were stained with CD125 (IL-5Rα) antibodies and with a suitable counterstaining. As a control, CD125 (IL-5Rα) 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 CD125 (IL-5Rα). Splenocytes from BALB/c mice were stained with CD125 (IL-5Rα) antibodies and with a suitable counterstaining. As a control, CD125 (IL-5Rα) 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 CD125 (IL-5Rα) (REA343). 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 CD125 (IL-5Rα) (REA343). 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 CD125 (IL-5Rα) (REA343). 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 CD125 (IL-5Rα) Antibody, anti-mouse, REAfinity™

Overview

Clone REA343 recognizes the mouse CD125 antigen, a single-pass type I membrane protein, also known as interleukin-5 receptor subunit α (IL-5Rα). The IL-5 receptor is a heterodimer composed of a ligand specific α subunit and a signal transducing β subunit which is shared by the receptors for IL-3, GM-CSF, and IL-5. CD125 is expressed on eosinophils, basophils, and B cells. IL-5 is a conserved hematopoietic cytokine which is mainly produced by Tʜ2 cells after stimulation with antigens, and by mast cells upon stimulation with an allergen/IgE complex or a calcium ionophore. The overexpression of IL-5
in vivo
significantly increases the numbers of eosinophils and B cells, while mice lacking the functional gene for IL-5 display various developmental and functional impairments in the B cell and eosinophil lineages.
Additional information: Clone REA343 displays negligible binding to Fc receptors.

Alternative names

CDw125, Il5r, IL-5RA

Detailed product information

Technical specifications

CloneREA343
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD125 (IL-5Rα)
Alternative names of antigenCDw125, Il5r, IL-5RA
Molecular mass of antigen [kDa]45
Distribution of antigenB cells, granulocytes, eosinophils, basophils
Entrez Gene ID16192
RRIDAB_2751542, AB_2819442, AB_2857775, AB_2654793, AB_2654796, AB_2654797, AB_2654798, AB_2654799, AB_2751556

References for CD125 (IL-5Rα) Antibody, anti-mouse, REAfinity™

Publications

  1. Takaki, S. et al. (1990) Molecular cloning and expression of the murine interleukin-5 receptor. EMBO J. 9(13): 4367-4374
  2. Mao, H. et al. (2006) Effect of interleukin-5 receptor-alpha short hairpin RNA-expressing vector on bone marrow eosinophilopoiesis in asthmatic mice. Adv. Ther. 23(6): 938-956
  3. Kusano, S. et al. (2012) Structural basis of interleukin-5 dimer recognition by its α receptor. Protein Sci. 21(6): 850-864

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