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      Alexa Fluor™ 647 Mouse Anti-Human SIT

      BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-Human SIT

      Clone SIT-01 (also known as SIT-01)

      (RUO)
      View all Formats
      Alexa Fluor™ 647 Mouse Anti-Human SIT
      Flow cytometric analysis of SIT expression in human peripheral blood lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD19 (Cat. No. 561741/555413). After staining, cells were treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to remove erythrocytes and fix leucocytes. The fixed leucocytes were permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723) and then stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732, Left Plot) or Alexa Fluor™ 647 SIT antibody (Cat. No. 568360/568367). The bivariate pseudocolor density plot showing the correlated expression of SIT (or Ig Isotype control staining) versus CD19 signals was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ software.
      Alexa Fluor™ 647 Mouse Anti-Human SIT
      Flow cytometric analysis of SIT expression in human peripheral blood lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD19 (Cat. No. 561741/555413). After staining, cells were treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to remove erythrocytes and fix leucocytes. The fixed leucocytes were permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723) and then stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732, Left Plot) or Alexa Fluor™ 647 SIT antibody (Cat. No. 568360/568367). The bivariate pseudocolor density plot showing the correlated expression of SIT (or Ig Isotype control staining) versus CD19 signals was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ software.
      Product Details
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      BD Pharmingen™
      SHP-2 interacting transmembrane adaptor protein; SIT; SIT1; signaling threshold-regulating transmembrane adapter 1
      Human (QC Testing)
      Mouse IgG1, κ
      Human Bacterially produced, recombinant, intracellular fragment of human SIT.
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      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 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 cell 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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      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. 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).
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. An isotype control should be used at the same concentration as the antibody of interest.
      8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      11. For U.S. patents that may apply, see bd.com/patents.
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      568360 Rev. 3
      Antibody Details
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      SIT-01

      The SIT-01 monoclonal antibody specifically recognizes an intracellular epitope of the transmembrane adaptor protein SIT1, also known as SIT, signaling-threshold regulating transmembrane adapter 1 and SHP2-interacting transmembrane adaptor protein. SIT1 is a disulfide-linked homodimeric glycoprotein with a substantial intracytoplasmic region that can be phosphorylated by Src- or Syk-family kinases and is expressed in lymphoid organs. This transmembrane adaptor protein plays a significant role in T cell development and peripheral T cell functions and is a key modulator of TCR-mediated signaling that regulates T-cell activation, homeostasis and tolerance. SIT1 has been proposed as an immunophenotypic marker in hematopathology.

      568360 Rev. 3
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      568360 Rev.3
      Citations & References
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      View product citations for antibody "568360" on CiteAb

      Development References (4)

      1. Hübener C, Mincheva A, Lichter P, Schraven B, Bruyns E. Complete sequence, genomic organization, and chromosomal localization of the human gene encoding the SHP2-interacting transmembrane adaptor protein (SIT).. Immunogenetics. 53(4):337-41. (Biology). View Reference
      2. Marie-Cardine A, Kirchgessner H, Bruyns E et al. SHP2-interacting transmembrane adaptor protein (SIT), a novel disulfide-linked dimer regulating human T cell activation. J Exp Med. 1999; 189(8):1181-1194. (Biology). View Reference
      3. Simeoni L, Posevitz V, Kölsch U, et al. The transmembrane adapter protein SIT regulates thymic development and peripheral T-cell functions.. Mol Cell Biol. 2005; 25(17):7557-68. (Biology: Flow cytometry). View Reference
      4. Tedoldi S, Paterson JC, Hansmann ML, et al. Transmembrane adaptor molecules: a new category of lymphoid-cell markers.. Blood. 2006; 107(1):213-21. (Biology). View Reference
      View All (4) View Less
      568360 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.