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      RB613 Mouse Anti-Stat3 (pY705)
      RB613 Mouse Anti-Stat3 (pY705)
      Analysis of Stat3 (pY705) expression in Human peripheral blood lymphocytes. Human whole blood was either left Unstimulated (dashed line histogram) or Stimulated (solid line histogram) with 100 ng/ml of BD Pharmingen™ Recombinant Human IL-6 protein (Cat. No. 550071) for 15 minutes at 37°C. The leukocytes were fixed and the erythrocytes were lysed in a single step by mixing and incubation with pre-warmed BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10 min at 37°C. The leukocytes were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), pelleted by centrifugation and then resuspended and left in BD Phosflow™ Perm Buffer III (Cat. No. 558052; chilled to -20°C before use) for 30 minutes on ice. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) and then stained with BD Phosflow™ RB613 Mouse Anti-Stat3 (pY705) antibody (Cat. No. 571242/571306). The fluorescence histograms showing Stat3 (pY705) expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.
      RB613 Mouse Anti-Stat3 (pY705)
      Analysis of Stat3 (pY705) expression in Human peripheral blood lymphocytes. Human whole blood was either left Unstimulated (dashed line histogram) or Stimulated (solid line histogram) with 100 ng/ml of BD Pharmingen™ Recombinant Human IL-6 protein (Cat. No. 550071) for 15 minutes at 37°C. The leukocytes were fixed and the erythrocytes were lysed in a single step by mixing and incubation with pre-warmed BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10 min at 37°C. The leukocytes were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656), pelleted by centrifugation and then resuspended and left in BD Phosflow™ Perm Buffer III (Cat. No. 558052; chilled to -20°C before use) for 30 minutes on ice. The cells were washed twice with BD Pharmingen™ Stain Buffer (FBS) and then stained with BD Phosflow™ RB613 Mouse Anti-Stat3 (pY705) antibody (Cat. No. 571242/571306). The fluorescence histograms showing Stat3 (pY705) expression were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.
      商品详情
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      BD Phosflow™
      Acute-phase response factor, APRF
      Human (QC Testing), Mouse (Tested in Development)
      Mouse IgG2a, κ
      Phosphorylated Human Stat3 Peptide
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      准备和存储

      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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      推荐的实验流程

      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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      商品通知

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. An isotype control should be used at the same concentration as the antibody of interest.
      7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      9. 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.
      10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      11. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
      12. CF™ is a trademark of Biotium, Inc.
      13. For U.S. patents that may apply, see bd.com/patents.
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      571242 Rev. 2
      抗体详情
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      4/P-STAT3

      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.  Seven Stat proteins have been cloned, each of which is differentially expressed and/or activated in a cytokine-specific and cell type-specific manner. 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. Stat3 activation occurs via tyrosine phosphorylation at Y705. Tyrosine phosphorylation in response to cytokine stimulation is generally mediated by JAK1. 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. In addition to tyrosine phosphorylation, Stat3 is also phosphorylated at S727 via the MAPK pathway.  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. Thus, phosphorylation of Y705 in Stat3 occurs in response to growth factors and cytokines, and is essential for normal transcription activity.

      The 4/P-STAT3 monoclonal antibody recognizes the phosphorylated Y705 of Stat3.

      571242 Rev. 2
      格式详情
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      RB613
      The BD Horizon RealBlue™ 613 (RB613) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492-nm and an emission maximum (Em Max) at 613-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB613 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with reduced excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB613 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-CF594. RB613 is on average brighter than PE-CF594 off the blue laser.
      altImg
      RB613
      Blue 488 nm
      492 nm
      613 nm
      571242 Rev.2
      报价单和参考
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      View product citations for antibody "571242" on CiteAb

      研发参考 (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). 查看参考
      2. Imada K, Leonard WJ. The Jak-STAT pathway. Mol Immunol. 2000; 37:1-11. (Biology). 查看参考
      3. Liu KD, Gaffen SL, Goldsmith MA. JAK/STAT signaling by cytokine receptors. Curr Opin Immunol. 1998; 10(3):271-278. (Biology). 查看参考
      4. Renner ED, Rylaarsdam S, Anover-Sombke S, et al. Novel signal transducer and activator of transcription 3 (STAT3) mutations, reduced T(H)17 cell numbers, and variably defective STAT3 phosphorylation in hyper-IgE syndrome. J Allergy Clin Immunol. 2008; 122(1):181-187. (Clone-specific: Flow cytometry). 查看参考
      5. 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). 查看参考
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      571242 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.