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      PE Mouse Anti-Human IFN-γ
      PE Mouse Anti-Human IFN-γ
      Flow cytometric analysis for IFN-γ in stimulated Rhesus macaque peripheral blood mononuclear cells (PBMC). PBMC from Rhesus macaque were stimulated for 6 hours with 50 ng/mL PMA (Sigma-Aldrich Cat. No. P-8139) and 500 ng/mL calcium ionophore A23187 (Sigma-Aldrich Cat. No. C-9275) in the presence of BD GolgiStop™ (Cat. No. 554724). Cells were then fixed and permeabilized using BD Cytofix/Cytoperm™ (Cat. No. 554714) followed by staining with FITC Mouse Anti-Human CD3 (Cat. No. 561807/555332/555916/561806) and PE Mouse Anti-Human IFN-γ antibody (Cat. No. 557074/554552/559326/561056). Two-color dot plot was derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
      PE Mouse Anti-Human IFN-γ
      Flow cytometric analysis for IFN-γ in stimulated Rhesus macaque peripheral blood mononuclear cells (PBMC). PBMC from Rhesus macaque were stimulated for 6 hours with 50 ng/mL PMA (Sigma-Aldrich Cat. No. P-8139) and 500 ng/mL calcium ionophore A23187 (Sigma-Aldrich Cat. No. C-9275) in the presence of BD GolgiStop™ (Cat. No. 554724). Cells were then fixed and permeabilized using BD Cytofix/Cytoperm™ (Cat. No. 554714) followed by staining with FITC Mouse Anti-Human CD3 (Cat. No. 561807/555332/555916/561806) and PE Mouse Anti-Human IFN-γ antibody (Cat. No. 557074/554552/559326/561056). Two-color dot plot was derived from gated events based on light scattering characteristics for lymphocytes. Flow cytometry was performed on a BD™ LSR II flow cytometry system.
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      BD Pharmingen™
      IFNG; Interferon-gamma; IFG; IFI; Type II interferon
      Rhesus, Cynomolgus, Baboon (QC Testing), Human (Tested in Development)
      Mouse BALB/c IgG1, κ
      Human IFN-γ from supernatants of S. aureus-stimulated PBMC
      Intracellular staining (flow cytometry) (Routinely Tested)
      20 µl
      3458
      AB_396572
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      557074 Rev. 7
      Antibody Details
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      4S.B3

      The 4S.B3 monoclonal antibody specifically binds to interferon-γ (IFN-γ). The immunogen used to generate this hybridoma was partially purified human IFN-γ obtained from supernatants of human PBMC stimulated with Staphylococcus aureus. Interferon-γ (IFN-γ) is a potent multifunctional cytokine that is produced by several activated cell types including NK, NKT, CD4+TCRαβ+, CD8+TCRαβ+, and TCRγδ+ T cells. IFN-γ exerts its biological effects through specific binding to the high-affinity IFN-γ Receptor Complex comprised of IFN-γRα (CD119) and IFN-γRβ subunits. In addition to its antiviral effects, IFN-γ upregulates a number of lymphoid cell functions including the antimicrobial and antitumor responses of macrophages, NK cells, and neutrophils. In addition, IFN-γ can exert strong regulatory influences on the proliferation, differentiation, and effector responses of B cell and T cell subsets. These influences can involve IFN-γ's capacity to boost MHC class I and II expression by antigen-presenting cells as well as to direct effects on B cells and T cells themselves. Human IFN-γ is a 14-18 kDa glycoprotein containing 143 amino acid residues.

      Clone 4S.B3 also cross-reacts with a cytoplasmic component of peripheral blood CD3+ lymphocytes of baboon, and both rhesus and cynomolgus macaque monkeys following five-hour treatment with phorbol myristic acetate (PMA) and Ca++ ionophore (A23187) in the presence of monensin. The staining pattern of 4S.B3 in CD3+ cells is similar to that observed with peripheral blood T lymphocytes from normal human donors. This reagent is useful for intracellular immunofluorescent staining for flow cytometric analysis to identify and enumerate IFN-γ + cells within a mixed cell population.

      557074 Rev. 7
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      557074 Rev.7
      Citations & References
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      View product citations for antibody "557074" on CiteAb

      Development References (4)

      1. Meager A. Characterization of interferons and immunoassays. In: Clemens MJ, Morris AG, Gearing AJH, ed. Lymphokines and Interferons. A Practical Approach. Oxford: IRL Press Ltd; 1987:105-127.
      2. 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). View Reference
      3. 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
      4. 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
      View All (4) View Less
      557074 Rev. 7

       

      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.