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Multicolor flow cytometric analysis of RORγt expression in Mouse thymocytes. C57BL/6 Mouse thymocytes were stained with BD OptiBuild™ BV421 Rat Anti-Mouse CD4 (Cat. No. 740007) and BD Horizon™ BUV395 Rat Anti-Mouse CD8a (Cat. No. 563786) antibodies, and then fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574). The fixed and permeabilized cells were then stained with either BD Horizon™ RB670 Mouse IgG2a, κ Isotype Control (Cat. No. 571958) or BD Horizon™ RB670 Mouse Anti-Mouse RORγt antibody (Cat. No. 571951/571952) at 0.125 µg/test. The overlapping histograms show the levels of either IgG2a Isotype Control (Left Panel) or 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 gated events with the forward and side light-scatter characteristics of intact cells. Samples were acquired using the BD FACSymphony™ A5 SE Cell Analyzer and were spectrally unmixed using FlowJo™ v10.10 Software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ RB670 Mouse Anti-Mouse RORγt
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- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
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- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
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- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
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配套商品
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.
研发参考 (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). 查看参考
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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). 查看参考
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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). 查看参考
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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). 查看参考
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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). 查看参考
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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: Intracellular Staining/Flow Cytometry). 查看参考
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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: Intracellular Staining/Flow Cytometry). 查看参考
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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: Intracellular Staining/Flow Cytometry). 查看参考
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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: Intracellular Staining/Flow Cytometry). 查看参考
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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: Intracellular Staining/Flow Cytometry). 查看参考
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