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      PE-Cy7 Mouse Anti-Human CD123 (IL-3 Receptor α)
      PE-Cy7 Mouse Anti-Human CD123 (IL-3 Receptor α)
      Multiparameter flow cytometric analysis of CD123 (IL-3 Receptor α) expression on Human peripheral blood leukocyte populations.  Human whole blood was stained with APC Mouse Anti-Human CD45RO (Cat No. 559865; Bottom Plots) and with either PE-Cy7 Mouse IgG1 κ Isotype Control (Cat. No. 565573; Left Plots) or PE-Cy7 Mouse Anti-Human CD123 (IL-3 Receptor α) antibody (Cat. No. 570777/570855; Right Plots) in the presence of BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (Cat. No. 570002). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat No. 349202).    Top Plots: 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 leukocytes.    Bottom Plots: Bivariate pseudocolor density plots showing the correlated expression of CD123 (IL-3 Receptor α) [or Ig Isotype control staining] versus CD45RO were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
      PE-Cy7 Mouse Anti-Human CD123 (IL-3 Receptor α)
      Multiparameter flow cytometric analysis of CD123 (IL-3 Receptor α) expression on Human peripheral blood leukocyte populations.  Human whole blood was stained with APC Mouse Anti-Human CD45RO (Cat No. 559865; Bottom Plots) and with either PE-Cy7 Mouse IgG1 κ Isotype Control (Cat. No. 565573; Left Plots) or PE-Cy7 Mouse Anti-Human CD123 (IL-3 Receptor α) antibody (Cat. No. 570777/570855; Right Plots) in the presence of BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (Cat. No. 570002). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat No. 349202).    Top Plots: 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 leukocytes.    Bottom Plots: Bivariate pseudocolor density plots showing the correlated expression of CD123 (IL-3 Receptor α) [or Ig Isotype control staining] versus CD45RO were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
      Product Details
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      BD Pharmingen™
      IL3RA; IL3R; IL-3R-alpha; IL-3RA
      Human (QC Testing)
      Mouse BALB/c IgG1, κ
      Human IL3RA Transfected Cell Line
      Flow cytometry (Routinely Tested)
      5 µl/test
      3563
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      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. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
      7. An isotype control should be used at the same concentration as the antibody of interest.
      8. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      11. Clones without a listed workshop have not been investigated in an HLDA workshop to receive a CD nomenclature. We use “CD” provisionally when our internal testing indicates that this clone binds to the same CD antigen as workshopped clones.
      12. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      13. For U.S. patents that may apply, see bd.com/patents.
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      570855 Rev. 1
      Antibody Details
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      6H6

      The 6H6 monoclonal antibody specifically recognizes the Interleukin-3 receptor alpha chain (IL-3Ra) which is also known as CD123. IL-3Ra (CD123) is a ~70 kDa type I transmembrane glycoprotein that is encoded by IL3RA (interleukin 3 receptor subunit alpha) which belongs to the type I cytokine receptor family within the immunoglobulin gene superfamily. This receptor chain consists of an extracellular region that contains an immunoglobulin-like N-terminal domain (NTD) with a fibronectin type III (FnIII) fold followed by two more FnIII domains that form the cytokine receptor module (CRM), a transmembrane region, and an intracellular tail. IL-3Ra (CD123) binds IL-3 specifically and with low affinity. IL-3Ra (CD123) forms a high-affinity signaling receptor for IL-3 (IL-3R) with the ß common chain (ßc; also known as, CD131) that is shared with the heterodimeric IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors. IL-3Ra (CD123) is variably expressed on certain hematopoietic progenitor cells, basophils, eosinophils, mast cells, monocytes, macrophages, dendritic cells, megakaryocytes, and on some B cells, endothelial cells, and leukemia cells. Bound IL-3 can signal through IL-3R to promote the activation, proliferation, differentiation, and viability of these cells. Amongst monoclonal antibodies specific for human IL-3Ra (CD123), the 6H6 and 9F5 antibodies do not block IL-3 binding to the IL-3R whereas the 7G3 antibody does block IL-3 binding to its receptor in a dose-dependent manner.

      570855 Rev. 1
      Format Details
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      PE-Cy7
      PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE-Cy7
      Yellow-Green 561 nm
      496 nm, 566 nm
      781 nm
      570855 Rev.1
      Citations & References
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      View product citations for antibody "570855" on CiteAb

      Development References (7)

      1. Broughton SE, Hercus TR, Hardy MP, et al. Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody.. Cell Rep. 2014; 8(2):410-9. (Biology). View Reference
      2. Kohrgruber N, Halanek N, Gröger M, et al. Survival, maturation, and function of CD11c- and CD11c+ peripheral blood dendritic cells are differentially regulated by cytokines.. J Immunol. 1999; 163(6):3250-9. (Clone-specific: Flow cytometry). 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. (Biology). View Reference
      4. Miyajima A. CDw123 (Interleukin 3 receptor α chain) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:854-855.
      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: Flow cytometry, Immunoprecipitation, Western blot). View Reference
      6. Yamada T, Sun Q, Zeibecoglou K, et al. IL-3, IL-5, granulocyte-macrophage colony-stimulating factor receptor alpha-subunit, and common beta-subunit expression by peripheral leukocytes and blood dendritic cells.. J Allergy Clin Immunol. 1998; 101(5):677-82. (Clone-specific: Flow cytometry). View Reference
      7. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
      View All (7) View Less
      570855 Rev. 1

      Please refer to Support Documents for Quality Certificates

       

      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.

      Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.