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RB545 Mouse Anti-Human CD123 (IL-3 Receptor α)
RB545 Mouse Anti-Human CD123 (IL-3 Receptor α)
Multiparameter flow cytometric analysis of CD123 (IL-3 Receptor α) expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ RB545 Mouse IgG2a, κ Isotype Control (Cat. No. 569277, Left Plot) or BD Horizon™ RB545 Mouse Anti-Human CD123 (IL-3 Receptor α) antibody (Cat. No. 569269/569270; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD123 (IL-3 Receptor α) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 Software.
Multiparameter flow cytometric analysis of CD123 (IL-3 Receptor α) expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ RB545 Mouse IgG2a, κ Isotype Control (Cat. No. 569277, Left Plot) or BD Horizon™ RB545 Mouse Anti-Human CD123 (IL-3 Receptor α) antibody (Cat. No. 569269/569270; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD123 (IL-3 Receptor α) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 Software.
Product Details
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BD Horizon™
IL3RA; IL-3RA; IL-3Rα; IL-3R-alpha; Interleukin-3 receptor subunit alpha
Human (QC Testing)
Mouse IgG2a, κ
Human IL-3Ra-transfected cells
Flow cytometry (Routinely Tested)
5 µl/test
3563
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  9. For U.S. patents that may apply, see bd.com/patents.
569270 Rev. 1
Antibody Details
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7G3

The 7G3 monoclonal antibody specifically recognizes human CD123, the 70 kDa IL-3 Receptor α (IL-3Rα) chain. CD123 associates with CD131, the 120-140 kDa Common β chain to form the IL-3 Receptor Complex. CD131 is shared with the receptors for interleukins IL-5 and GM-CSF. IL-3Rα is expressed on hematopoietic progenitors and plays an important role in hematopoietic progenitor cell growth and differentiation. It is also expressed by mast cells, macrophages and a CD5+ B cell subset. This antibody has been reported to block the binding of 125I-IL-3 to high and low affinity IL-3 receptors. In functional experiments, this antibody was found to inhibit acute myeloid leukemia cell proliferation, basophil histamine release, endothelial cell-mediated IL-8 secretion, and neutrophil transmigration. This antibody has been reported to be useful for immunoprecipitation, Western blot and immunofluorescent staining for flow cytometry. At the Fifth HLDA Workshop, the human IL-3 receptor was designated CD123.

569270 Rev. 1
Format Details
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RB545
The BD Horizon RealBlue™ 545 (RB545) fluorochrome is part of the BD family of blue (488-nm) laser dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 496-nm and an emission maximum (Em Max) at 545-nm. Driven by BD innovation, RB545 can be used on spectral cytometers and is designed to be excited by the blue (488-nm) laser with minimal excitation by the 561-nm yellow-green laser. RB545 has minimal spillover into yellow-green detectors and a brightness level similar to FITC.  Given its unique emission max, on a spectral instrument RB545 can be used simultaneously with FITC and PE to provide an additional color off of the blue laser. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RB545
Blue 488 nm
496 nm
545 nm
569270 Rev.1
Citations & References
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Development References (7)

  1. Du W, Li XE, Sipple J, Pang Q. Overexpression of IL-3Rα on CD34+CD38- stem cells defines leukemia-initiating cells in Fanconi anemia AML.. Blood. 2011; 117(16):4243-52. (Clone-specific: Flow cytometry, Inhibition, Neutralization). View Reference
  2. Korpelainen EI, Gamble JR, Smith WB, et al. The receptor for interleukin 3 is selectively induced in human endothelial cells by tumor necrosis factor alpha and potentiates interleukin 8 secretion and neutrophil transmigration.. Proc Natl Acad Sci USA. 1993; 90(23):11137-41. (Biology). View Reference
  3. Macardle PJ, Chen Z, Shih CY, et al. Characterization of human leucocytes bearing the IL-3 receptor. Cell Immunol. 1996; 168(1):59-68. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). View Reference
  4. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
  5. Smith WB, Guida L, Sun Q, et al. Neutrophils activated by granulocyte-macrophage colony-stimulating factor express receptors for interleukin-3 which mediate class II expression. Blood. 1995; 86(10):3938-3944. (Clone-specific: Flow cytometry, Functional assay, Inhibition). View Reference
  6. Sun Q, Woodcock JM, Rapoport A, et al. Monoclonal antibody 7G3 recognizes the N-terminal domain of the human interleukin-3 (IL-3) receptor alpha-chain and functions as a specific IL-3 receptor antagonist.. Blood. 1996; 87(1):83-92. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Neutralization). View Reference
  7. Williams BA, Wang XH, Leyton JV, et al. CD16+NK-92 and anti-CD123 monoclonal antibody prolongs survival in primary human acute myeloid leukemia xenografted mice.. Haematologica. 2018; 103(10):1720-1729. (Clone-specific: Cell separation, Flow cytometry). View Reference
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569270 Rev. 1

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