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PE-Cy7 Mouse Anti-Mouse RORγt
PE-Cy7 Mouse Anti-Mouse RORγt

Multicolor flow cytometric analysis of RORɣt expression in mouse thymocytes. BALB/c mouse thymocytes were stained with BD Horizon™ BUV395 Rat Anti-Mouse CD4 (Cat. No. 740208) and BD Horizon™ BV510 Rat Anti-Mouse CD8a (Cat. No. 563068) antibodies, then fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). The fixed and permeabilized cells were then stained with either PE-Cy7 Mouse IgG2a, κ Isotype Control (Cat. No. 552868; Left Plot) or PE-Cy7 Mouse Anti-Mouse RORγt antibody (Cat. No. 567305; Right Plot) at 0.03 µg/test. The overlapping histograms show the levels of IgG2a Isotype Control or RORɣt staining in CD4+ CD8- single-positive (dashed line histogram) or CD4+ CD8+ double-positive (solid line histogram) thymocytes. The fluorescence histograms showing RORɣt expression (or Ig isotype control staining) 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™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Multicolor flow cytometric analysis of RORɣt expression in mouse thymocytes. BALB/c mouse thymocytes were stained with BD Horizon™ BUV395 Rat Anti-Mouse CD4 (Cat. No. 740208) and BD Horizon™ BV510 Rat Anti-Mouse CD8a (Cat. No. 563068) antibodies, then fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). The fixed and permeabilized cells were then stained with either PE-Cy7 Mouse IgG2a, κ Isotype Control (Cat. No. 552868; Left Plot) or PE-Cy7 Mouse Anti-Mouse RORγt antibody (Cat. No. 567305; Right Plot) at 0.03 µg/test. The overlapping histograms show the levels of IgG2a Isotype Control or RORɣt staining in CD4+ CD8- single-positive (dashed line histogram) or CD4+ CD8+ double-positive (solid line histogram) thymocytes. The fluorescence histograms showing RORɣt expression (or Ig isotype control staining) 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™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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
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 unconjugated antibody and free dye were 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 CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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. 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.
  4. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. 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 PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  9. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567305 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.

567305 Rev. 1
Format Details
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PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-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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PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
567305 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
567305 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

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

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.