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Flow cytometric analysis of CD278 expression on stimulated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were cultured with Phytohemagglutinin (PHA; Sigma L-1668) for 3 days. The stimulated cells were then stained with either BD Horizon™ BUV395 Mouse IgG1 κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon BUV395 Mouse Anti-Human CD278 antibody (Cat. No. 564777; solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Horizon™ BUV395 Mouse Anti-Human CD278
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The DX29 monoclonal antibody specifically binds to human CD278, which is also known as Inducible Costimulator (ICOS) or Inducible T-cell Costimulator. ICOS is a homodimeric type I transmembrane glycoprotein with an approximate molecular weight of 50-60 kDa. It is a member of the CD28 family and is highly expressed on activated T cells. CD278 is the receptor for ICOS-ligand (also known as, CD275, B7-H2, B7RP-1, or LICOS). Like CD28, ICOS can provide a costimulatory signal for T cell activation, proliferation and cytokine production. It is not expressed on resting or activated B cells, monocytes, NK cells, granulocytes, dendritic cells or platelets. Unlike the constitutively expressed CD28, ICOS is de novo expressed upon cellular activation. Reports describe similarities between CD28 and ICOS in T cell activation, such as the costimulation of cytokine production. However, it has been suggested that ICOS may play a greater role in IL-10 production. In the presence of IL-10, purified recombinant human ICOS protein significantly increased in vitro B cell growth stimulated by pokeweed mitogen (PWM) and enhanced production of IgG.
The antibody was conjugated to BD Horizon BUV395 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is optimal for multicolor flow cytometry because it has little to no spillover into other detectors. With an Ex Max at 348 nm and an Em Max at 395 nm, BD Horizon BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.
Development References (7)
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Aicher A, Hayden-Ledbetter M, Brady WA, et al. Characterization of human inducible costimulator ligand expression and function. J Immunol. 2000; 164(9):4689-4696. (Biology). View Reference
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Dong C, Nurieva RI. Regulation of immune and autoimmune responses by ICOS. J Autoimmun. 2003; 21(3):255-260. (Biology). View Reference
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Fos C, Salles A, Lang V, et al. ICOS ligation recruits the p50alpha PI3K regulatory subunit to the immunological synapse. J Immunol. 2008; 181(3):1969-1977. (Clone-specific: Flow cytometry). View Reference
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Kallinich T, Beier KC, Gelfand EW, Kroczek RA, Hamelmann E. Co-stimulatory molecules as potential targets for therapeutic intervention in allergic airway disease. Clin Exp Allergy. 2005; 35(12):1521-1534. (Biology). View Reference
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Okamoto N, Tezuka K, Kato M, Abe R, Tsuji T. PI3-kinase and MAP-kinase signaling cascades in AILIM/ICOS- and CD28-costimulated T-cells have distinct functions between cell proliferation and IL-10 production. Biochem Biophys Res Commun. 2003; 310(3):691-702. (Biology). View Reference
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Sakamoto S, Tezuka K, Tsuji T, Hori N, Tamatani T. AILIM/ICOS: its expression and functional analysis with monoclonal antibodies. Hybrid Hybridomics. 2001; 20(5-6):293-303. (Biology). View Reference
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Witsch EJ, Peiser M, Hutloff A, et al. ICOS and CD28 reversely regulate IL-10 on re-activation of human effector T cells with mature dendritic cells. Eur J Immunol. 2002; 32(9):2680-2686. (Biology). View Reference
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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.