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      PerCP-Cy™5.5 Mouse Anti-Stat6
      Product Details
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      BD Phosflow™
      STAT6; Signal transducer and activator of transcription 6; IL-4-STAT
      Human (QC Testing), Mouse, Rat (Tested in Development)
      Mouse IgG1
      Human Stat6 C-terminal Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_2738622
      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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
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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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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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. An isotype control should be used at the same concentration as the antibody of interest.
      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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      6. 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.
      7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. PE-Cy5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the PE-Cy5 tandem fluorochrome, extra care must be taken when using dual-laser cytometers which may directly excite both PE and Cy5™.
      10. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
      11. Cy is a trademark of GE Healthcare.
      12. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      13. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564149 Rev. 2
      Antibody Details
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      23/Stat6

      STATs (signal transducers and activators of transcription) are critical mediators of the biologic activity of cytokines including Interleukins (IL) 2-5, IL-7, IL-15, GM-CSF, erythropoietin and growth hormone.  Ligand-receptor interaction leads to activation of constitutively associated JAK family kinases and subsequent recruitment/activation of STATs by tyrosine phosphorylation.  Active STATs 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.  Stat6 plays an important role in signaling pathways that lead to the differentiation of T helper type 2 (Th2) cells from uncommitted CD4 T cell precursors.  Moreover, IL-4, secreted by activated T lymphocytes, basophils, and mast cells, induces specific gene expression via the induction of tyrosine phosphorylation of Stat6 at tyrosine 641 (Y641).  The SH3:SH2 domain of Stat6 associates with tyrosine-phosphorylated IL-4 receptor and the proximal Jak kinase phosphorylates Stat6 at Y641 on the C-terminal side of the SH2 domain.  Stat6 is then released from the receptor, dimerizes, and is thought to contact the basal transcription machinery by binding to p300/CBP.  While Stat6 is widely expressed in human tissues, it exhibits elevated expression in peripheral blood lymphocytes, colon, intestine, ovary, prostate, thymus, spleen, kidney, liver, lung, and placenta.

      The 23/Stat6 monoclonal antibody recognizes Stat6, regardless of phosphorylation status.

      564149 Rev. 2
      Format Details
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      PerCP-Cy5.5
      PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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
      PerCP-Cy5.5
      Blue 488 nm
      482 nm
      676 nm
      564149 Rev.2
      Citations & References
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      Development References (10)

      1. Bartoli M, Gu X, Tsai NT, et al. Vascular endothelial growth factor activates STAT proteins in aortic endothelial cells. J Biol Chem. 2000; 275(43):33189-33192. (Clone-specific: Immunofluorescence, Immunoprecipitation, Western blot). View Reference
      2. 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
      3. Dent AL, Hu-Li J, Paul WE, Staudt LM. T helper type 2 inflammatory disease in the absence of interleukin 4 and transcription factor STAT6. Proc Natl Acad Sci U S A. 1998; 95(23):13823-13828. (Biology). View Reference
      4. Heim MH. The Jak-STAT pathway: specific signal transduction from the cell membrane to the nucleus. Eur J Clin Invest. 1996; 26(1):1-12. (Clone-specific). View Reference
      5. Hou J, Schindler U, Henzel WJ, Ho TC, Brasseur M, McKnight SL. An interleukin-4-induced transcription factor: IL-4 Stat. Science. 1994; 265(5179):1701-1706. (Biology).
      6. Mikita T, Campbell D, Wu P, Williamson K, Schindler U. Requirements for interleukin-4-induced gene expression and functional characterization of Stat6. Mol Cell Biol. 1996; 16(10):5811-5820. (Biology).
      7. Peine M, Rausch S, Helmstetter C, et al. Stable T-bet(+)GATA-3(+) Th1/Th2 hybrid cells arise in vivo, can develop directly from naive precursors, and limit immunopathologic inflammation. PLoS ONE. 2013; 11(8):e10016331. (Clone-specific: Flow cytometry). View Reference
      8. Quelle FW, Shimoda K, Thierfelder W, et al. Cloning of murine Stat6 and human Stat6, Stat proteins that are tyrosine phosphorylated in responses to IL-4 and IL-3 but are not required for mitogenesis. Mol Cell Biol. 1995; 15(6):3336-3343. (Biology).
      9. Waite KJ, Floyd ZE, Arbour-Reily P, Stephens JM. Interferon-gamma-induced regulation of peroxisome proliferator-activated receptor gamma and STATs in adipocytes. J Biol Chem. 2001; 276(10):7062-7068. (Clone-specific: Western blot). View Reference
      10. Xia Z, Salzler RR, Kunz DP, et al. A novel serine-dependent proteolytic activity is responsible for truncated signal transducer and activator of transcription proteins in acute myeloid leukemia blasts. Cancer Res. 2001; 61(4):1747-1753. (Clone-specific: Western blot). View Reference
      View All (10) View Less
      564149 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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