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Alexa Fluor® 647 Mouse Anti-Human RORγt
Alexa Fluor® 647 Mouse Anti-Human RORγt

Flow cytometric analysis of human RORγt expression in human SR cells. Cells from the human SR cell line (ATCC CRL-2262) were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and then stained with either Alexa Fluor® 647 Mouse IgG2b, κ Isotype Control (Cat. No. 558713; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human RORγt antibody (Cat. No. 563620; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of human RORγt expression in human SR cells. Cells from the human SR cell line (ATCC CRL-2262) were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and then stained with either Alexa Fluor® 647 Mouse IgG2b, κ Isotype Control (Cat. No. 558713; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human RORγt antibody (Cat. No. 563620; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
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BD Pharmingen™
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
AB_2738324
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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. 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).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
563620 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.

563620 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
563620 Rev.1
Citations & References
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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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563620 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.