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PE Mouse Anti-Human CD123
PE Mouse Anti-Human CD123
Multiparameter flow cytometric analysis of CD123 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 554648; Left Plot) or PE Mouse Anti-Human CD123 antibody (Cat. No. 554529/561058; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Solution (Cat. No. 555899). Bivariate pseudocolor density plots showing the correlated expression of CD123 (IL-3 Receptor α) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis was performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
PE Mouse Anti-Human CD123
Peripheral blood lymphocytes analyzed on a FACScan (BDIS, San Jose, CA)
Multiparameter flow cytometric analysis of CD123 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG2a, κ Isotype Control (Cat. No. 554648; Left Plot) or PE Mouse Anti-Human CD123 antibody (Cat. No. 554529/561058; Right Plot) at 0.5 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Solution (Cat. No. 555899). Bivariate pseudocolor density plots showing the correlated expression of CD123 (IL-3 Receptor α) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis was performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Peripheral blood lymphocytes analyzed on a FACScan (BDIS, San Jose, CA)
Product Details
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BD Pharmingen™
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)
0.2 mg/ml
3563
AB_395457
Aqueous buffered solution containing protein stabilizer and ≤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 unconjugated antibody and free dye were 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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
561058 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.

561058 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
561058 Rev.1
Citations & References
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Development References (6)

  1. 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
  2. Macardle PJ, Chen Z, Shih CY, et al. Characterization of human leucocytes bearing the IL-3 receptor. Cell Immunol. 1996; 168(1):59-68. (Biology). View Reference
  3. 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.
  4. 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
  5. 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, Immunoprecipitation, Neutralization, Western blot). View Reference
  6. Zola H. Detection of cytokine receptors by flow cytometry. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: Green Publishing Associates and Wiley-Interscience; 1995:6.21.1-6.21.18.
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561058 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.