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R718 Mouse Anti-Mouse RORγt
R718 Mouse Anti-Mouse RORγt
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.
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.
Product Details
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BD Horizon™
RORγT; RORgt; RORgamma t; RORgammaT; Rorc2; Rorg; TOR; Thor; Nr1f3
Mouse (QC Testing)
Mouse IgG2a, κ
Mouse RORγt Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567362 Rev. 1
Antibody Details
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Q31-378

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.

567362 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567362 Rev.1
Citations & References
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View product citations for antibody "567362" on CiteAb

Development References (10)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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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567362 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.