Skip to main content Skip to navigation
PE-CF594 Mouse Anti-Human RORγt
PE-CF594 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™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; dashed line histogram) or BD Horizon™ PE-CF594 Mouse Anti-Human RORγt antibody (Cat. No. 567532; 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™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; dashed line histogram) or BD Horizon™ PE-CF594 Mouse Anti-Human RORγt antibody (Cat. No. 567532; 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
Down Arrow Up Arrow


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 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 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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  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. 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).
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. CF™ is a trademark of Biotium, Inc.
  9. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  10. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. 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.
  11. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  12. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  13. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
567532 Rev. 1
Antibody Details
Down Arrow Up Arrow
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.

This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg, 610/20-nm filter).

567532 Rev. 1
Format Details
Down Arrow Up Arrow
PE-CF594
BD Horizon™ PE-CF594 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 with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-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 green (532-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.
altImg
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
567532 Rev.1
Citations & References
Down Arrow Up Arrow

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
View All (6) View Less
567532 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.