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R718 Mouse Anti-Human RORγt
R718 Mouse Anti-Human RORγt

Flow cytometric analysis of human RORγt expression in human SR cells. Cells from the human SR (Pleural effusion lymphoblast, ATCC CRL-2262) cell line were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and then stained with either BD Horizon™ R718 Mouse IgG2b, κ Isotype Control (Cat. No. 566952; dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human RORγt antibody (Cat. No. 567097; solid line histogram). The fluorescence histogram showing the expressed levels of RORγt (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

Flow cytometric analysis of human RORγt expression in human SR cells. Cells from the human SR (Pleural effusion lymphoblast, ATCC CRL-2262) cell line were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and then stained with either BD Horizon™ R718 Mouse IgG2b, κ Isotype Control (Cat. No. 566952; dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human RORγt antibody (Cat. No. 567097; solid line histogram). The fluorescence histogram showing the expressed levels of RORγt (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.

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
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. 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).
  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.
567097 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™ Red 718, which has been developed exclusively by 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.

567097 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
567097 Rev.1
Citations & References
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View product citations for antibody "567097" on CiteAb

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
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567097 Rev. 1

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