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      V450 Rat Anti-Human IL-6
      V450 Rat Anti-Human IL-6
      Flow cytometric analysis of IL-6 expression by stimulated human monocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 6 hours with lipopolysaccharide (LPS; 100 ng/ml final concentration) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing monensin) (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with PE Mouse Anti-Human CD14 monoclonal antibody (Cat. No. 555398), fixed and permeabilized using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ Buffer (Cat. No. 554723), and stained with BD Horizon™ V450 Rat Anti-Human IL-6 antibody (Cat. No. 561446, solid line histogram) or a BD Horizon™ V450 Rat IgG1, κ Isotype Control (Cat No. 560535, dashed line histogram). The fluorescence histograms were derived from CD14-positive events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      V450 Rat Anti-Human IL-6
      Flow cytometric analysis of IL-6 expression by stimulated human monocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 6 hours with lipopolysaccharide (LPS; 100 ng/ml final concentration) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing monensin) (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with PE Mouse Anti-Human CD14 monoclonal antibody (Cat. No. 555398), fixed and permeabilized using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ Buffer (Cat. No. 554723), and stained with BD Horizon™ V450 Rat Anti-Human IL-6 antibody (Cat. No. 561446, solid line histogram) or a BD Horizon™ V450 Rat IgG1, κ Isotype Control (Cat No. 560535, dashed line histogram). The fluorescence histograms were derived from CD14-positive events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Horizon™
      IL6; Interleukin-6; BSF-2; CDF; HGF; HSF; IFNB2
      Human (QC Testing)
      Rat IgG1
      Human IL-6 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_10681055
      Aqueous buffered solution containing protein stabilizer 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™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.
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      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. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
      4. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
      5. 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.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      561446 Rev. 1
      Antibody Details
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      MQ2-13A5

      The MQ2-13A5 monoclonal antibody specifically binds to human interleukin-6 (IL-6). IL-6 is a multifunctional cytokine that plays a central role in host defense mechanisms, including hematopoiesis, immune responses (eg, T and B cell activation and differentiation) and acute phase reactions. IL-6 can be expressed by a variety of cells including monocytes/macrophages, eosinophils, fibroblasts, vascular endothelial cells, bone marrow stromal cells, mesangial cells, hepatocytes, keratinocytes, astrocytes, T lymphocytes, B lymphocytes, and various tumor cells. IL-6 production is upregulated by cells in response to bacterial products such as lipopolysaccharide, viruses and other pro-inflammatory cytokines such as IL-1, TNF , and IFN-γ. IL-6 transcription is downregulated by cells in response to IL-4 and IL-10. The functional IL-6 Receptor (IL-6R) complex consists of two transmembrane glycoproteins, an 80-kDa low-affinity ligand-binding receptor subunit (IL-6Rα/CD126) and a 130 kDa (gp130/CD130) subunit that binds to IL-6-IL-6Rα to form the high-affinity signal transducing complex. Abnormal expression of IL-6 is related to the pathogenesis of many diseases including neoplastic (eg, multiple myeloma) and autoimmune diseases (eg, rheumatoid arthritis). The immunogen used to generate this hybridoma was COS-7 -expressed recombinant human IL-6.

      The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

      561446 Rev. 1
      Format Details
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      V450
      BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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      V450
      Violet 405 nm
      405 nm
      450 nm
      561446 Rev.1
      Citations & References
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      Development References (4)

      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. (Biology: ELISA, Neutralization). 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. (Biology: ELISA, Neutralization). View Reference
      3. Gaines Das RE, Poole S. The international standard for interleukin-6. Evaluation in an international collaborative study. J Immunol Methods. 1993; 160(2):147-153. (Biology: ELISA, Neutralization). View Reference
      4. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: IC/FCM Block). View Reference
      View All (4) View Less
      561446 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.

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