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BV421 Mouse Anti-Mouse RORγt
BV421 Mouse Anti-Mouse RORγt

Multicolor flow cytometric analysis of RORγt expression in mouse thymocytes. BALB/c mouse thymocytes were stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091) and FITC Rat Anti-Mouse CD8a (Cat. No. 553031/553030/561966) 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™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439) or BD Horizon™ BV421 Mouse Anti-Mouse RORγt antibody (Cat. No. 562894). 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 APC Rat Anti-Mouse CD4 (Cat. No. 553051/561091) and FITC Rat Anti-Mouse CD8a (Cat. No. 553031/553030/561966) 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™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439) or BD Horizon™ BV421 Mouse Anti-Mouse RORγt antibody (Cat. No. 562894). 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 Horizon™
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_2687545
Aqueous buffered solution containing BSA and ≤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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

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. Brilliant Violet™ 421 is a trademark of Sirigen.
  4. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562894 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 Brilliant Violet™ 421 and has been developed in collaboration with Sirigen. With an Ex Max of 407-nm and Em Max at 421-nm, Brilliant Violet™ 421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). Brilliant Violet™ 421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562894 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562894 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
562894 Rev. 1

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