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      BV650 Mouse Anti-Human CD116
      BV650 Mouse Anti-Human CD116
      Two-parameter flow cytometric analysis of CD116 expression on human peripheral blood leucocytes. Human peripheral blood was treated with PharmLyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes prior to staining. The leucocytes were then preincubated with purified normal human polyclonal IgG (5 μg/10^6 cells) and stained with either BD Horizon™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; Left Panel) or BD Horizon BV650 Mouse Anti-Human CD116 antibody (Cat No. 564044; Right Panel). Two parameter flow cytometric contour plots showing the correlated expression of CD116 (or Ig Isotype control staining) versus side-light scatter (SSC) signals were derived from events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      BV650 Mouse Anti-Human CD116
      Two-parameter flow cytometric analysis of CD116 expression on human peripheral blood leucocytes. Human peripheral blood was treated with PharmLyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes prior to staining. The leucocytes were then preincubated with purified normal human polyclonal IgG (5 μg/10^6 cells) and stained with either BD Horizon™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; Left Panel) or BD Horizon BV650 Mouse Anti-Human CD116 antibody (Cat No. 564044; Right Panel). Two parameter flow cytometric contour plots showing the correlated expression of CD116 (or Ig Isotype control staining) versus side-light scatter (SSC) signals were derived from events with the forward and side light-scatter characteristics of viable leucocyte populations. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Horizon™
      CSF2RA; GM-CSF Receptor alpha; GM-CSFRα; GMCSFRA; GMR, SMDP4
      Human (QC Testing)
      Mouse IgG1, κ
      Recombinant human GM-CSFR
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      V C007
      AB_2738560
      Aqueous buffered solution containing BSA 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 with BD Horizon™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 were removed.
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      Recommended Assay Procedures

      Note: Certain human cell lines or cell types (e.g., neutrophils, monocytes) can first be treated with reagents that block receptors for the Fc regions of immunoglobulin to avoid nonspecific immunofluorescent staining mediated by Fc receptors.

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349).

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      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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. 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.
      5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
      8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564044 Rev. 2
      Antibody Details
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      hGMCSFR-M1

      The hGMCSFR-M1 antibody reacts with the subunit (GM-CSFR) of the human Granulocyte-Macrophage Colony-Stimulating Factor Receptor complex. This 75-85 kD subunit is also known as CD116. The hGMCSFR-M1 antibody was first clustered at the Fifth International Workshop on Human Leucocyte Differentiation Antigens. The GM-CSFR subunit associates with the 120-140 kD βc subunit (common subunit, CD131), that is shared with the receptors for interleukins IL-3 and IL-5. Both of the chains of the GM-CSFR complex are involved in ligand binding and intracellular signaling. The α chain appears to transmit most of the biological signals. CD116 is expressed by a variety of myeloid cell lines, hematopoietic and non-hematopoetic tumor cells, and normal cell types including monocytes, macrophages, neutrophils, eosinophils, myeloid dendritic cells, endothelial cells, fibroblasts, and placental trophoblasts. Lymphocytes are negative for GM-CSFR expression. Reports suggest that GM-CSFR plays a role in myeloid lineage growth and differentiation. The immunogen used to generate the hGMCSFR-M1 hybridoma was recombinant human GM-CSFR.

      The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.

      564044 Rev. 2
      Format Details
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      BV650
      The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-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.
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      BV650
      Violet 405 nm
      406 nm
      649 nm
      564044 Rev.2
      Citations & References
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      Development References (12)

      1. Böhm I, Speck U, Schild HH. Pilot study on basophil activation induced by contrast medium.. Fundam Clin Pharmacol. 2011; 25(2):267-76. (Biology). View Reference
      2. Cheng YX, Foster B, Holland SM, et al. CD2 identifies a monocyte subpopulation with immunoglobulin E-dependent, high-level expression of Fc epsilon RI.. Clin Exp Allergy. 2006; 36(11):1436-45. (Biology). View Reference
      3. Eder M, Ernst TJ, Ganser A, et al. low affinity chimeric human alpha/beta-granulocyte-macrophage colony-stimulating factor receptor induces ligand-dependent proliferation in a murine cell line. J Biol Chem. 1994 ; 269(48):30173-30180. (Biology). View Reference
      4. Foster B, Metcalfe DD, Prussin C. Human dendritic cell 1 and dendritic cell 2 subsets express FcepsilonRI: correlation with serum IgE and allergic asthma.. J Allergy Clin Immunol. 2003; 112(6):1132-8. (Biology). View Reference
      5. Jokhi PP, King A, Jubinsky PT, Loke YW. Demonstration of the low affinity alpha subunit of the granulocyte-macrophage colony-stimulating factor receptor (GM-CSF-R alpha) on human trophoblast and uterine cells. J Reprod Immunol. 1994; 26(2):147-164. (Biology). View Reference
      6. Jubinsky PT, Laurie AS, Nathan DG, Yetz-Aldepe J, Sieff CA. Expression and function of the human granulocyte-macrophage colony-stimulating factor receptor alpha subunit. Blood. 1994; 84(12):4174-4185. (Biology). View Reference
      7. Kubista B, Trieb K, Herbacek I, Micksche. Effect of granulocyte-macrophage colony-stimulating factor on natural-killer cell mediated cytotoxicity. Int J Biochem Cell Biol. 2003; 35(7):1056-1060. (Clone-specific: Flow cytometry). View Reference
      8. Kubota T, Mukai K, Minegishi Y, Karasuyama H. Different stabilities of the structurally related receptors for IgE and IgG on the cell surface are determined by length of the stalk region in their alpha-chains.. J Immunol. 2006; 176(11):7008-14. (Biology). View Reference
      9. Ronco LV, Silverman SL, Wong SG, Slamon DJ, Park LS, Gasson JC. Identification of conserved amino acids in the human granulocyte-macrophage colony-stimulating factor receptor alpha subunit critical for function. Evidence for formation of a heterodimeric receptor complex prior to ligand binding. J Biol Chem. 1994; 269(1):277-283. (Biology). View Reference
      10. 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.
      11. Stacchini A, Fubini L, Aglietta M. Flow cytometric detection and quantitative analysis of the GM-CSF receptor in human granulocytes and comparison with the radioligand binding assay. Cytometry. 1996; 24(4):374-381. (Clone-specific: Flow cytometry). View Reference
      12. Wognum AW, Westerman Y, Visser TP, Wagemaker G. Distribution of receptors for granulocyte-macrophage colony-stimulating factor on immature CD34+ bone marrow cells, differentiating monomyeloid progenitors, and mature blood cell subsets. Blood. 1994; 84(3):764-774. (Biology). View Reference
      View All (12) View Less
      564044 Rev. 2

      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.

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