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Alexa Fluor® 647 Mouse Anti-Human CD116
Alexa Fluor® 647 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 Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human CD116 antibody (Cat No. 564046). 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.
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 Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714) or Alexa Fluor® 647 Mouse Anti-Human CD116 antibody (Cat No. 564046). 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 Pharmingen™
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_2738562
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

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.

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. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564046 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.

564046 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564046 Rev.2
Citations & References
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Development References (9)

  1. 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
  2. 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
  3. 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
  4. 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
  5. Lanza F, Moretti S, Papa S, Malavasi F, Castoldi G. Report on the Fifth International Workshop on Human Leukocyte Differentiation Antigens, Boston, November 3-7, 1993.. Haematologica. 79(4):374-86. (Biology). View Reference
  6. 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
  7. 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.
  8. 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
  9. 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
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564046 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.