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PerCP-Cy™5.5 Mouse Anti-Mouse RORγt
PerCP-Cy™5.5 Mouse Anti-Mouse RORγt

Multicolor flow cytometric analysis of RORγt expression in mouse thymocytes. BALB/c mouse thymocytes were stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835) and PE Rat Anti-Mouse CD8a (Cat. No. 553033/553032/561095) 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 PerCP-Cy™5.5 Mouse IgG2a, κ Isotype Control (Cat. No. 550927) or PerCP-Cy™5.5 Mouse Anti-Mouse RORγt antibody (Cat. No. 562683). The overlapping histograms show the levels of IgG2a Isotype Control (Left Panel) and RORγt (Right Panel) staining in CD4+ CD8- single-positive (dashed line histogram) or CD4+ CD8+ double-positive (solid line histogram) thymocytes. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.

Multicolor flow cytometric analysis of RORγt expression in mouse thymocytes. BALB/c mouse thymocytes were stained with FITC Rat Anti-Mouse CD4 (Cat. No. 553047/553046/561835) and PE Rat Anti-Mouse CD8a (Cat. No. 553033/553032/561095) 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 PerCP-Cy™5.5 Mouse IgG2a, κ Isotype Control (Cat. No. 550927) or PerCP-Cy™5.5 Mouse Anti-Mouse RORγt antibody (Cat. No. 562683). The overlapping histograms show the levels of IgG2a Isotype Control (Left Panel) and RORγt (Right Panel) staining in CD4+ CD8- single-positive (dashed line histogram) or CD4+ CD8+ double-positive (solid line histogram) thymocytes. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.

Product Details
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BD Pharmingen™
RORγT; RORgt; RORgamma t; RORgammaT; Rorc2; Rorg; TOR; Thor; Nr1f3
Mouse (QC Testing)
Mouse IgG2a, κ
Mouse RORγt Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2737720
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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Recommended Assay Procedures

Optimal staining occurs when using the BD Pharmingen™ PerCP-Cy™5.5 Mouse Anti-Mouse RORγt antibody with cells that have been fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. 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.
  7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  8. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
562683 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.

562683 Rev. 1
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
562683 Rev.1
Citations & References
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Development References (9)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. Medvedev A, Yan ZH, Hirose T, Giguere V, Jetten AM. Cloning of a cDNA encoding the murine orphan receptor RZR/ROR gamma and characterization of its response element. Gene. 1996; 181(1-2):199-206. (Biology). View Reference
  9. Unutmaz D. RORC2: the master of human Th17 cell programming. Eur J Immunol. 2009; 39(6):1452-1455. (Biology). View Reference
View All (9) View Less
562683 Rev. 1

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