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Alexa Fluor® 647 Rat Anti-Human GM-CSF
Alexa Fluor® 647 Rat Anti-Human GM-CSF
Multicolor analysis of GM-CSF expressed by HiCK-2 cells. HiCK-2 (Human intracellular CytoKine-2) Cytokine Positive Control Cells (Cat. No. 555062) were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat No. 557906, Left Panel) or Alexa Fluor® 647 Mouse anti-Human GM-CSF antibody (Cat No. 562257, Right Panel) by using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the expressed levels of GM-CSF (or Ig isotype control staining) versus cellular autofluorescence (measured in the FL1 channel) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor analysis of GM-CSF expressed by HiCK-2 cells. HiCK-2 (Human intracellular CytoKine-2) Cytokine Positive Control Cells (Cat. No. 555062) were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with either Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat No. 557906, Left Panel) or Alexa Fluor® 647 Mouse anti-Human GM-CSF antibody (Cat No. 562257, Right Panel) by using BD Biosciences Intracellular Cytokine Staining protocol. Two-color flow cytometric dot plots showing the expressed levels of GM-CSF (or Ig isotype control staining) versus cellular autofluorescence (measured in the FL1 channel) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
CSF2; Colony stimulating factor 2 (granulocyte-macrophage); CSF; GMCSF
Human (QC Testing)
Rat LEW, also known as Lewis IgG2a
Recombinant human GM-CSF
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_11152076
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. 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).
  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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
562257 Rev. 1
Antibody Details
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BVD2-21C11

The BVD2-21C11 monoclonal antibody specifically binds to human Granulocyte/Macrophage - Colony Stimulating Factor (GM-CSF). Human GM-CSF is encoded by the CSF2 gene and is also known as Colony Stimulating Factor 2.  GM-CSF is produced by activated T lymphocytes, macrophages, endothelial cells, fibroblasts, stromal cells and other cell types including B lymphocytes, mast cells, eosinophils, and osteoblasts.  GM-CSF stimulates the survival, proliferation and/or differentiation of various cell types including neutrophils, eosinophils, macrophages, dendritic cells, megakaryocytes, erythroid cells, endothelial cells and their precursors. The immunogen used to generate the BVD2-21C11 hybridoma was recombinant human GM-CSF. The BVD2-21C11 antibody has been reported to crossreact with GM-CSF from the rhesus monkey. BVD2-21C11 is a neutralizing antibody.

The binding of conjugated BVD2-21C11 antibody has been shown to be blocked by preincubation with recombinant human GM-CSF (0.1 μg; Cat. No. 550068) and by preincubation of the fixed/permeabilized cells with unlabeled BVD2-21C11 antibody (Cat. No. 554503) prior to staining.  Please view the PE Rat anti-Human GM-CSF (Cat. No. 554507) Technical Data Sheet for additional data.

562257 Rev. 1
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
562257 Rev.1
Citations & References
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Development References (5)

  1. Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA). View Reference
  2. Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: ELISA, Immunoprecipitation, Neutralization). View Reference
  3. Abrams JS, Silver JE, Van Dyke RE, Gleich GI. Eosinophil-active cytokines in human disease: development and use of monoclonal antibodies to IL-3, IL-5 and GMCSF. In: Gleich GJ and Kay AB, ed. Eosinophils in Allergy and Inflammation. New York: Dekker; 1994:133-157.
  4. Bacchetta R, de Waal Malefijt R, Yssel H. Host-reactive CD4+ and CD8+ T cell clones isolated from a human chimera produce IL-5, IL-2, IFN-gamma and granulocyte/macrophage-colony-stimulating factor but not IL-4. J Immunol. 1990; 144(3):902-908. (Clone-specific: ELISA, Neutralization). View Reference
  5. Kita H, Ohnishi T, Okubo Y, Weiler D, Abrams JS, Gleich GJ. Granulocyte/macrophage colony-stimulating factor and interleukin 3 release from human peripheral blood eosinophils and neutrophils. J Exp Med. 1991; 174(3):745-748. (Clone-specific: ELISA, Neutralization). View Reference
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562257 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.