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    BV650 Mouse Anti-Human RORγt
    BV650 Mouse Anti-Human RORγt
    Flow cytometric analysis of human RORγt expression in human SR cells. Cells from the human SR cell line (ATCC CRL-2262) were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and then stained with either BD Horizon™ BV650 Mouse IgG2b, κ Isotype Control (Cat. No. 563437; dashed line histogram) or BD Horizon™ BV650 Mouse Anti-Human RORγt antibody (Cat. No. 563424; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
    BV650 Mouse Anti-Human RORγt
    Flow cytometric analysis of human RORγt expression in human SR cells. Cells from the human SR cell line (ATCC CRL-2262) were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and then stained with either BD Horizon™ BV650 Mouse IgG2b, κ Isotype Control (Cat. No. 563437; dashed line histogram) or BD Horizon™ BV650 Mouse Anti-Human RORγt antibody (Cat. No. 563424; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
    Product Details
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    BD Horizon™
    RORgt; ROR gamma t; RORC; NR1F3; RORG; RZRG; TOR; RZR-GAMMA
    Human (QC Testing)
    Mouse IgG2b, κ
    Human RORγt Recombinant Protein
    Intracellular staining (flow cytometry) (Routinely Tested)
    5 µl
    AB_2738197
    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™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 were 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. 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. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
    6. Brilliant Violet™ 650 is a trademark of Sirigen.
    7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
    8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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    563424 Rev. 1
    Antibody Details
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    Q21-559

    The Q21-559 monoclonal antibody recognizes ROR gamma t (RORγt), an isoform of ROR gamma (RORγ). RORγt is a DNA-binding transcription factor that is encoded by the RORC (RAR-related orphan receptor C) gene. It belongs to the ROR/RZR orphan nuclear receptor family. Two protein isoforms are produced from the RORC gene by alternative splicing: RORγ or RORγt. The RORγ isoform is expressed in many tissues, including thymus, lung, liver, kidney, muscle, and brown fat. However the tissue distribution of the second isoform RORγt is highly restricted to the thymus where it is expressed exclusively in immature CD4+CD8+ thymocytes and in peripheral Th17 and LTi cells. RORγt reportedly functions as an essential transcription factor for the development of secondary lymphoid tissues. It also plays an important regulatory role in thymopoiesis and effector T cell differentiation, especially for Th17 cells.

    The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. This dye is a tandem fluorochrome of BD Horizon™ BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm.  BD Horizon™ BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter).  Due to the excitation and emission characteristics of the acceptor dye, there will be  spillover into the APC and Alexa Fluor® 700 detectors.  However, the spillover can be corrected through compensation as with any other dye combination.

    563424 Rev. 1
    Format Details
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    BV650
    The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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    BV650
    Violet 405 nm
    406 nm
    649 nm
    563424 Rev.1
    Citations & References
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    Development References (6)

    1. Acosta-Rodriguez EV, Rivino L, Geginat J, et al. Surface phenotype and antigenic specificity of human interleukin 17-producing T helper memory cells. Nat Immunol. 2007; 8(6):639-646. (Biology). View Reference
    2. Bechtel S, Rosenfelder H, Duda A, et al. The full-ORF clone resource of the German cDNA Consortium. BMC Biochem. 2007; 8:399-410. (Biology). View Reference
    3. Crellin NK, Trifari S, Kaplan CD, Cupedo T, Spits H. Human NKp44+IL-22+ cells and LTi-like cells constitute a stable RORC+ lineage distinct from conventional natural killer cells. J Exp Med. 2010; 207(2):281-290. (Biology). View Reference
    4. Hirose T, Smith RJ, Jetten AM. ROR gamma: the third member of ROR/RZR orphan receptor subfamily that is highly expressed in skeletal muscle. Biochim Biophys Acta. 1994; 205(3):1976-1983. (Biology). View Reference
    5. Jetten AM. Retinoid-related orphan receptors (RORs): critical roles in development, immunity, circadian rhythm, and cellular metabolism. Nucl Recept Signal. 2009; 7:1-32. (Biology). View Reference
    6. Zielinski CE, Mele F, Aschenbrenner D, et al. Pathogen-induced human TH17 cells produce IFN-gamma or IL-10 and are regulated by IL-1beta. Nature. 2012; 484(7395):514-518. (Biology). View Reference
    View All (6) View Less
    563424 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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