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      BV421 Mouse Anti-Human SIT
      BV421 Mouse Anti-Human SIT
      Two-color flow cytometric analysis of SIT expression in Human peripheral blood lymphocytes.  Human whole blood was stained with BD Horizon™ BB515 Mouse Anti-Human CD19 (Cat. No. 564456).  After staining, cells were treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to remove erythrocytes and fix leukocytes. The fixed leukocytes were permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723) and then stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438, Left Plot) or BD Horizon™ BV421 SIT antibody (Cat. No. 569799/569873). 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.
      BV421 Mouse Anti-Human SIT
      Two-color flow cytometric analysis of SIT expression in Human peripheral blood lymphocytes.  Human whole blood was stained with BD Horizon™ BB515 Mouse Anti-Human CD19 (Cat. No. 564456).  After staining, cells were treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to remove erythrocytes and fix leukocytes. The fixed leukocytes were permeabilized using BD Perm/Wash™ Buffer (Cat. No. 554723) and then stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438, Left Plot) or BD Horizon™ BV421 SIT antibody (Cat. No. 569799/569873). 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 Horizon™
      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/test
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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 unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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.
      3. 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).
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. An isotype control should be used at the same concentration as the antibody of interest.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. 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.
      8. For U.S. patents that may apply, see bd.com/patents.
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      569799 Rev. 1
      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.

      569799 Rev. 1
      Format Details
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      BV421
      The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV421
      Violet 405 nm
      407 nm
      423 nm
      569799 Rev.1
      Citations & References
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      View product citations for antibody "569799" 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
      569799 Rev. 1

      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.