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Multicolor flow cytometric analysis of RORγt expression in mouse thymocytes. BALB/c mouse thymocytes were stained with PE Rat Anti-Mouse CD4 (Cat. No. 553048/553049/561837) and BD Horizon™ BV510 Rat Anti-Mouse CD8a (Cat. No. 563068) antibodies, and then fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The fixed and permeabilized cells were then stained with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 566949) or BD Horizon™ R718 Mouse Anti-Mouse RORγt antibody (Cat. No. 567362) 0.06 ug/test. The overlapping histograms show the levels of IgG2a Isotype Control (Left Panel) and RORγt (Right Panel) staining in CD4+ CD8- single-positive (dashed line histograms) or CD4+ CD8+ double-positive (solid line histograms) thymocytes. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ R718 Mouse Anti-Mouse RORγt
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Preparation And Storage
Recommended Assay Procedures
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The Q31-378 monoclonal antibody recognizes mouse RORgamma t (RORγt), an isoform of RORgamma (RORγ). RORγt is a DNA-binding transcription factor that belongs to the ROR/RZR orphan nuclear receptor family. RORγt is expressed exclusively by lymphoid cells including CD4+CD8+ thymocytes, peripheral CD4+ Th17 and CD8+ Tc17 cells, NKT cells and innate lymphoid cells such as lymphoid tissue inducer (LTi) cells. RORγt plays essential roles in thymopoiesis, T cell homeostasis, differentiation of effector T lymphocytes and the development of secondary lymphoid tissues including lymph nodes and Peyer's patches.
The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor® 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor® 700.
Development References (10)
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Eberl G, Littman DR. The role of the nuclear hormone receptor RORgt in the development of lymph nodes and Peyer's patches. Immunol Rev. 2003; 195(195):81-90. (Biology). View Reference
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Eberl G, Marmon S, Sunshine MJ, Rennert PD, Choi Y, Littman DR. An essential function for the nuclear receptor RORgamma(t) in the generation of fetal lymphoid tissue inducer cells. Nat Immunol. 2004; 5(1):64-73. (Biology). View Reference
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Hamada H, Garcia-Hernandez MdlL, Reome JB, et al. Tc17, a unique subset of CD8 T cells that can protect against lethal influenza challenge. J Immunol. 2009; 182(6):3469-3481. (Biology). View Reference
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He YW, Deftos ML, Ojala EW, Bevan MJ. RORgamma t, a novel isoform of an orphan receptor, negatively regulates Fas ligand expression and IL-2 production in T cells. Immunity. 1998; 9(6):797-806. (Biology). View Reference
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Ivanov, II, McKenzie BS, Zhou L, et al. The orphan nuclear receptor RORgammat directs the differentiation program of proinflammatory IL-17+ T helper cells. Cell. 2006; 126(6):1121-1133. (Biology). View Reference
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Lee JY, Hall JA, Kroehling L, et al. Serum Amyloid A Proteins Induce Pathogenic Th17 Cells and Promote Inflammatory Disease. Cell. 2020; 180(1):79-91.e16. (Clone-specific: Flow cytometry). View Reference
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Metidji A, Omenetti S, Crotta S, et al. The Environmental Sensor AHR Protects from Inflammatory Damage by Maintaining Intestinal Stem Cell Homeostasis and Barrier Integrity.. Immunity. 2018; 49(2):353-362.e5. (Clone-specific: Flow cytometry). View Reference
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Sidwell T, Liao Y, Garnham AL, et al. Attenuation of TCR-induced transcription by Bach2 controls regulatory T cell differentiation and homeostasis. Nature communications. 2020; 11(1):252. (Clone-specific: Flow cytometry). View Reference
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Wang H, Hogquist KA. CCR7 defines a precursor for murine iNKT cells in thymus and periphery. eLife. 2018; 7:e34793. (Clone-specific: Flow cytometry). View Reference
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Wei SC, Sharma R, Anang NAS, et al. Negative Co-stimulation Constrains T Cell Differentiation by Imposing Boundaries on Possible Cell States. Immunity. 2019; 50(4):1084-1098.e1010. (Clone-specific: Flow cytometry). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.