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      BUV395 Mouse Anti-Human TNF
      BUV395 Mouse Anti-Human TNF
      Flow cytometric analysis of TNF expression in stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 6 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of  BD GolgiStop™  Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ BUV395 Mouse IgG1 κ Isotype Control (Cat. No. 563547; Left Panel) or BD Horizon™ BUV395 Mouse Anti-Human TNF antibody (Cat. No. 563996; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of TNF (or Ig Isotype control staining) versus Autofluorescence for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      BUV395 Mouse Anti-Human TNF
      Flow cytometric analysis of TNF expression in stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 6 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of  BD GolgiStop™  Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), and fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either BD Horizon™ BUV395 Mouse IgG1 κ Isotype Control (Cat. No. 563547; Left Panel) or BD Horizon™ BUV395 Mouse Anti-Human TNF antibody (Cat. No. 563996; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of TNF (or Ig Isotype control staining) versus Autofluorescence for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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      BD Horizon™
      Tumor necrosis factor alpha; TNF-a; TNF-α; TNFSF2; Cachectin
      Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
      Mouse IgG1, κ
      Recombinant Human TNF
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_2738533
      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 BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 were removed.
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      推奨アッセイ手順

      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. 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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 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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      563996 Rev. 2
      抗体の詳細
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      MAb11

      The MAb11 monoclonal antibody specifically binds to human tumor necrosis factor (TNF, also known as TNF-α) protein. TNF is an efficient juxtacrine, paracrine and endocrine mediator of inflammatory and immune functions. It regulates the growth and differentiation of a variety of cell types. TNF is cytotoxic for transformed cells when in conjunction with IFN-γ. It is secreted by activated monocytes/macrophages and other cells such as B cells, T cells and fibroblasts. The immunogen used to generate the MAb11 hybridoma was recombinant human TNF. The MAb11 antibody has been reported to crossreact with Rhesus Macaque TNF.

      The antibody was conjugated to BD Horizon™ BUV395 which has been exclusively developed by BD Biosciences as an optimal dye for use on a 355 nm laser equipped instrument. With an Ex Max at 348 nm  and an Em Max at 395 nm, this dye has virtually no spillover into any other detector. BD Horizon™ BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.

      563996 Rev. 2
      フォーマットの詳細
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      BUV395
      The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BUV395
      Ultraviolet 355 nm
      348 nm
      395 nm
      563996 Rev.2
      引用＆参考文献
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      Development References (10)

      1. Black RA, Rauch CT, Kozlosky CJ, et al. A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. Nature. 1997; 385(6618):729-733. (Biology). View Reference
      2. Danis VA, Franic GM, Rathjen DA, Brooks PM. Effects of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-2, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and IL-6 on the production of immunoreactive IL-1 and TNF-alpha by human monocytes. Clin Exp Immunol. 1991; 85(1):143-150. (Clone-specific: ELISA). View Reference
      3. Jason J, Larned J. Single-cell cytokine profiles in normal humans: comparison of flow cytometric reagents and stimulation protocols. J Immunol Methods. 1997; 207(1):13-22. (Clone-specific: Flow cytometry). View Reference
      4. Kriegler M, Perez C, DeFay K, Albert I, Lu SD. A novel form of TNF/cachectin is a cell surface cytotoxic transmembrane protein: ramifications for the complex physiology of TNF. Cell. 1988; 53(1):45-53. (Biology). View Reference
      5. 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
      6. Rathjen DA, Cowan K, Furphy LJ, Aston R. Antigenic structure of human tumour necrosis factor: recognition of distinct regions of TNF alpha by different tumour cell receptors. Mol Immunol. 1991; 28(1-2):79-86. (Clone-specific: ELISA). View Reference
      7. Sander B, Andersson J, Andersson U. Assessment of cytokines by immunofluorescence and the paraformaldehyde-saponin procedure. Immunol Rev. 1991; 119:65-93. (Biology). View Reference
      8. Sopper S, Stahl-Hennig C, Demuth M, Johnston IC, Dorries R, ter Meulen V. Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. Cytometry. 1997; 29(4):351-362. (Biology). View Reference
      9. Verdier F, Aujoulat M, Condevaux F, Descotes J. Determination of lymphocyte subsets and cytokine levels in cynomolgus monkeys. Toxicology. 1995; 105(1):81-90. (Biology). View Reference
      10. Wang AM, Creasey AA, Ladner MB, et al. Molecular cloning of the complementary DNA for human tumor necrosis factor. Science. 1985; 228(4696):149-154. (Biology). View Reference
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      563996 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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