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      RY586 Mouse Anti-Stat1 (pY701)
      RY586 Mouse Anti-Stat1 (pY701)
      Two-color flow cytometric analysis of Stat1 (pY701) expression in Human CD4+ T lymphocytes. Untreated (dashed line histogram) or Treated (solid line histogram) Human peripheral blood mononuclear cells (PBMC) from the BD Phosflow™ T Cell Kit Lyophilized Cells (Catalog No. 560760) were reconstituted in BD Pharmingen™ Stain Buffer (Cat. No. 554656) for 15 minutes at room temperature. These cells are fixed, permeabilized, and lyophilized Human PBMC that are either Untreated or Treated with stimulators [including Phorbol 12-Myristate 13-Acetate (PMA), and Recombinant Human Interferon-alpha (IFN-α), Interleukin-2 (IL-2), IL-4, and IL-6 proteins]. The reconstituted cells were then washed and stained with Alexa Fluor™ 488 Mouse Anti-Human CD4 antibody (Cat. No. 557695) and BD Phosflow™ RY586 Mouse Anti-Stat1 (pY701) antibody (Cat. No. 568528/568529). Histograms showing Stat1 (pY701) expression were derived from CD4-positive gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD™ LSR II Flow Cytometry System and FlowJo™ software.
      RY586 Mouse Anti-Stat1 (pY701)
      Two-color flow cytometric analysis of Stat1 (pY701) expression in Human CD4+ T lymphocytes. Untreated (dashed line histogram) or Treated (solid line histogram) Human peripheral blood mononuclear cells (PBMC) from the BD Phosflow™ T Cell Kit Lyophilized Cells (Catalog No. 560760) were reconstituted in BD Pharmingen™ Stain Buffer (Cat. No. 554656) for 15 minutes at room temperature. These cells are fixed, permeabilized, and lyophilized Human PBMC that are either Untreated or Treated with stimulators [including Phorbol 12-Myristate 13-Acetate (PMA), and Recombinant Human Interferon-alpha (IFN-α), Interleukin-2 (IL-2), IL-4, and IL-6 proteins]. The reconstituted cells were then washed and stained with Alexa Fluor™ 488 Mouse Anti-Human CD4 antibody (Cat. No. 557695) and BD Phosflow™ RY586 Mouse Anti-Stat1 (pY701) antibody (Cat. No. 568528/568529). Histograms showing Stat1 (pY701) expression were derived from CD4-positive gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD™ LSR II Flow Cytometry System and FlowJo™ software.
      Product Details
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      BD Phosflow™
      Signal Transducer and Activator of Transcription-1; STAT91; ISGF-3; CANDF7
      Human (QC Testing), Mouse (Tested in Development), Rat (Predicted)
      Mouse IgG2a
      Phosphorylated Human Stat1 Peptide
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      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 unreacted dye was removed.
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      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.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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).
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. 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.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
      8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      9. CF™ is a trademark of Biotium, Inc.
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      568528 Rev. 1
      Antibody Details
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      4a

      Stat (Signal transducer and activators of transcription) proteins mediate the biological activity of cytokines, including interleukins, interferons, erythropoietin, and growth factors.  Ligand-receptor interaction activates constitutively-associated JAK family kinases as well as subsequent recruitment and activation of Stat proteins by tyrosine phosphorylation.  Active Stat proteins then move to the nucleus to promote transcription of cytokine-inducible genes.  Seven Stat proteins have been cloned, each of which is differentially expressed and/or activated in a cytokine-specific and cell type-specific manner.  Stat1 and Stat2 are components of the ISGF3 (Interferon-Stimulated Gene Factor 3) complex, the primary transcription activator induced by interferon binding to a specific cell-surface receptor.  Stat1 has two alternatively spliced isoforms, 91-kDa Stat1α and 84-kDa Stat1β; Stat1α has 38 additional C-terminal amino acids.  In response to the binding of IFNα, IFNγ, EGF, PDGF, or CSF-1 to their respective receptors, the Stat1 subunits become tyrosine-phosphorylated at Y701, and the complex translocates to the nucleus. This forms an active complex that includes the DNA-binding p48 subunit, and is responsible for modulating interferon-stimulated genes (ISGs) transciption.

      The 4a monoclonal antibody recognizes the phosphorylated Y701 in Stat1α and Stat1β.

      568528 Rev. 1
      Format Details
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      RY586
      The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
      altImg
      RY586
      Yellow-Green 561 nm
      564 nm
      586 nm
      568528 Rev.1
      Citations & References
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      View product citations for antibody "568528" on CiteAb

      Development References (6)

      1. Bromberg J, Darnell JE. The role of STATs in transcriptional control and their impact on cellular function. Oncogene. 2000; 19(21):2468-2473. (Biology). View Reference
      2. Darnell JE Jr. STATs and gene regulation. Science. 1997; 277(5332):1630-1635. (Biology). View Reference
      3. Fu XY, Zhang JJ. Transcription factor p91 interacts with the epidermal growth factor receptor and mediates activation of the c-fos gene promoter. Cell. 1993; 74(6):1135-1145. (Biology). View Reference
      4. Perez OD, Mitchell D, Campos R, Gao GJ, Li L, Nolan GP. Multiparameter analysis of intracellular phosphoepitopes in immunophenotyped cell populations by flow cytometry. Curr Protoc Cytom. 2005; 6.20.1-6.20.22. (Clone-specific: Flow cytometry). View Reference
      5. Suni MA, Maino VC. Flow cytometric analysis of cell signaling proteins. Methods Mol Biol. 2011; 717:155-169. (Clone-specific). View Reference
      6. Tanaka S, Saito Y, Kunisawa J, et al. Development of mature and functional human myeloid subsets in hematopoietic stem cell-engrafted NOD/SCID/IL2rgammaKO mice. J Immunol. 2012; 188(12):6145-6155. (Clone-specific: Flow cytometry). View Reference
      View All (6) View Less
      568528 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.