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      PE Mouse anti-Stat3
      PE Mouse anti-Stat3
      Analysis of Stat3 in human peripheral blood lymphocytes.  Human peripheral blood mononuclear cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes at 37ºC, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 559320, shaded histogram) or PE Mouse anti-Stat3 (open histogram). For data analysis, lymphocytes were selected by their scatter profile. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
      PE Mouse anti-Stat3
      Analysis of Stat3 in human epithelioid carcinoma.  HeLa S3 cells (ATCC CCL 2.2) were either transfected with Stat3 RNAi (shaded histogram) or untreated (open histogram).  The cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with PE Mouse anti-Stat3. Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.
      PE Mouse anti-Stat3 PE Mouse anti-Stat3
      Analysis of Stat3 in human peripheral blood lymphocytes.  Human peripheral blood mononuclear cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes at 37ºC, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 559320, shaded histogram) or PE Mouse anti-Stat3 (open histogram). For data analysis, lymphocytes were selected by their scatter profile. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
      Analysis of Stat3 in human epithelioid carcinoma.  HeLa S3 cells (ATCC CCL 2.2) were either transfected with Stat3 RNAi (shaded histogram) or untreated (open histogram).  The cells were fixed (BD Cytofix™ Fixation Buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with PE Mouse anti-Stat3. Flow cytometry was performed on a BD FACSArray™ bioanalyzer system.
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      BD Phosflow™
      STAT3; Acute-phase response factor; APRF; HIES
      Human (QC Testing), Mouse, Rat (Predicted)
      Mouse BALB/c IgG1, λ
      Human Stat3 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      20 µl
      AB_1645535
      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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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      Recommended Assay Procedures

      This antibody conjugate is suitable for intracellular staining of human cell lines and peripheral blood mononuclear cells using BD Cytofix™ Fixation Buffer.  Either BD Phosflow™ Perm Buffer II or Perm Buffer III may be used.  We have observed that BD Phosflow™ Perm/Wash Buffer I gives unsatisfactory results with this antibody.

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      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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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      560391 Rev. 2
      Antibody Details
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      M59-50

      Stat (Signal transducer and activators of transcription) proteins are critical mediators of the biologic activity of cytokines, including interleukins, interferons, erythropoietin, and growth factors.  Ligand-receptor interaction leads to activation of constitutively associated JAK family kinases and subsequent recruitment/activation of Stat proteins by tyrosine phosphorylation.  Active Stat proteins then move to the nucleus to promote transcription of cytokine-inducible genes.  Stat3 is a 92-kDa protein that is activated as a DNA-binding protein through cytokines, such as IL-6, and growth factors, such as EGF. Upon activation, Stat3 dimerizes, translocates to the nucleus and binds DNA response elements, thereby regulating gene expression. It has been reported that Stat3 binds to DNA as a homodimer, but it is also capable of binding as a heterodimer with Stat1. Stat3 is widely expressed and can bind to the sis-inducible element (SIE) site from the c-fos promoter. This site is similar to the GAS element that is present in IFN-γ induced genes.

      The M59-50 monoclonal antibody recognizes Stat3 (isoform 1) regardless of phosphorylation status. The specificity of this antibody conjugate for flow cytometric analysis was validated by confirming that RNA-mediated interference (RNAi) of the specific protein reduced the staining of the cells (see figure).  Furthermore, the capacity of the RNAi to down-regulate the expression of the relevant protein was confirmed by western blot analysis.

      560391 Rev. 2
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE 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 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      560391 Rev.2
      Citations & References
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      Development References (5)

      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. Kanai M, Konda Y, Nakajima T, et al . Differentiation-inducing factor-1 (DIF-1) inhibits STAT3 activity involved in gastric cancer cell proliferation via MEK-ERK-dependent pathway. Oncogene. 2003; 22(22):548-554. (Biology). View Reference
      4. Kirito K, Osawa M, Morita H. A functional role of Stat3 in in vivo megakaryopoiesis. Blood. 2002; 99(9):3220-3227. (Biology). View Reference
      5. Smith PD, Crompton MR. Expression of v-src in mammary epithelial cells induces transcription via STAT3. Biochem J. 1998; 15:331-381. (Biology). View Reference
      View All (5) View Less
      560391 Rev. 2

      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.

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