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Purified Mouse Anti-Human CD123
Purified Mouse Anti-Human CD123

Multiparameter flow cytometric analysis of CD123 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either Purified Mouse IgG2a, κ Isotype Control (Cat. No. 555571; Left Plot) or Purified Mouse Anti-Human CD123 antibody (Cat. No. 554527; Right Plot) at 0.5 µg/test, followed by Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 553999) and PE Streptavidin (Cat. No. 554061). 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.

Multiparameter flow cytometric analysis of CD123 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either Purified Mouse IgG2a, κ Isotype Control (Cat. No. 555571; Left Plot) or Purified Mouse Anti-Human CD123 antibody (Cat. No. 554527; Right Plot) at 0.5 µg/test, followed by Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 553999) and PE Streptavidin (Cat. No. 554061). 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.

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), Blocking, Immunoprecipitation, Neutralization, Western blot (Reported)
0.5 mg/ml
3563
AB_395455
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

Flow cytometry: This antibody has been tested by immunofluorescent staining (≤ 1 µg/million cells) with flow cytometric analysis to assure specificity and reactivity. For flow cytometric applications, a three step labeling procedure is recommended for amplifying signal.

Blocking: This antibody has been found to block the binding of 125I-IL-3 to high and low affinity IL-3 receptors and can neutralize IL-3 bioactivity. For neutralization studies we recommend our no sodium azide and low endotoxin (NA/LE) format (Cat. No. 554526).

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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
554527 Rev. 2
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.

554527 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
554527 Rev.2
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. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence). 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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554527 Rev. 2

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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.