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      Alexa Fluor® 488 Mouse Anti-Btk (pY551)/Itk (pY511)
      Alexa Fluor® 488 Mouse Anti-Btk (pY551)/Itk (pY511)
      Analysis of Btk (pY551) in activated human B lymphoma cells.  Ramos cells (ATCC CRL-1596) were serum starved overnight and then either stimulated with 5 mM hydrogen peroxide for 15 minutes (shaded histogram) or unstimulated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 488 anti-Btk (pY551)/Itk (pY511). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
      Alexa Fluor® 488 Mouse Anti-Btk (pY551)/Itk (pY511)
      Analysis of Btk (pY551) in activated human B lymphoma cells.  Ramos cells (ATCC CRL-1596) were serum starved overnight and then either stimulated with 5 mM hydrogen peroxide for 15 minutes (shaded histogram) or unstimulated (open histogram).  The cells were fixed (BD Cytofix™ buffer, Cat. No. 554655) for 10 minutes, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for at least 30 minutes, and then stained with Alexa Fluor® 488 anti-Btk (pY551)/Itk (pY511). Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
      商品详情
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      BD Phosflow™
      Human (QC Testing)
      Mouse IgG1, κ
      Phosphorylated Human Btk Peptide
      Intracellular staining (flow cytometry) (Routinely Tested)
      20 µl
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      准备和存储

      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® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
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      商品通知

      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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      3. 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.
      4. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
      5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      6. 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.
      7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      558130 Rev. 3
      抗体详情
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      24a/BTK (Y551)

      Bruton's tyrosine kinase (Btk) is a nonreceptor tyrosine kinase whose function is critical for proper B cell development and signaling.  The activity of Btk is regulated by Src mediated phosphorylation of the kinase domain at tyrosine 551 (Y551).  This event induces Btk kinase activity and subsequent autophosphorylation at Y223.  Phosphorylated Btk then associates with the cell membrane via the interaction of the PH domain with phosphatidylinositol 3, 4, 5-triphosphate.  

      The Tec family kinase Itk plays a critical role in signal transduction downstream of the T cell antigen receptor and has been implicated in the activation of phospholipase C-γ1 (PLCγ1), a key regulator of calcium mobilization and extracellular signal-regulated kinase (ERK) activation.  Itk is regulated by an activating transphosphorylation event in which Y511 in the kinase domain is phosphorylated by Lck.  

      The 24a/BTK (Y551) monoclonal antibody recognizes the Y551-phosphorylated form of human Btk and the Y511 phosphorylated form of human Itk.  

      558130 Rev. 3
      格式详情
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      Alexa Fluor™ 488
      Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 488
      Blue 488 nm
      494 nm
      517 nm
      558130 Rev.3
      报价单和参考
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      研发参考 (3)

      1. Mahajan S, Fargnoli J, Burkhardt AL, Kut SA, Saouaf SJ, Bolen JB. Src family protein tyrosine kinases induce autoactivation of Bruton's tyrosine kinase. Mol Cell Biol. 1995; 15:5304-5311. (Biology). 查看参考
      2. Rawlings DJ, Scharenberg AM, Park H, et al. Activation of BTK by a phosphorylation mechanism initiated by SRC family kinases. Science. 1996; 271:822-825. (Biology). 查看参考
      3. Wilcox HM, Berg LJ.. Itk phosphorylation sites are required for functional activity in primary T cells. J Biol Chem. 2003; 278:37112-37121. (Biology).
      558130 Rev. 3

      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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