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      RY610 Rat Anti-Mouse CD4
      RY610 Rat Anti-Mouse CD4
      Two-color flow cytometric analysis of CD4 expression on Mouse splenic leukocytes. BALB/c Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/32 antibody (Mouse Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody (Cat. No. 563565) and with either BD Horizon™ RY610 Rat IgG2b, κ Isotype Control (Cat. No. 571153; Left Plot) or BD Horizon™ RY610 Rat Anti-Mouse CD4 antibody (Cat. No. 571146/571147; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable splenic leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      RY610 Rat Anti-Mouse CD4
      Two-color flow cytometric analysis of CD4 expression on Mouse splenic leukocytes. BALB/c Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/32 antibody (Mouse Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BUV395 Hamster Anti-Mouse CD3e antibody (Cat. No. 563565) and with either BD Horizon™ RY610 Rat IgG2b, κ Isotype Control (Cat. No. 571153; Left Plot) or BD Horizon™ RY610 Rat Anti-Mouse CD4 antibody (Cat. No. 571146/571147; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3e was derived from gated events with the forward and side light-scatter characteristics of viable splenic leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Horizon™
      Cd4; CD4 antigen; L3T4; Ly-4; T-cell surface antigen T4/Leu-3
      Mouse (QC Testing)
      Rat LEW, also known as Lewis IgG2b, κ
      Mouse CTL clone V4
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      12504
      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 unreacted dye was 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.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. 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.
      5. 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.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      8. CF™ is a trademark of Biotium, Inc.
      9. For U.S. patents that may apply, see bd.com/patents.
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      571146 Rev. 1
      Antibody Details
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      GK1.5

      The GK1.5 monoclonal antibody specifically binds to the mouse CD4 (L3T4) differentiation antigen. CD4 is expressed on most thymocytes, a subpopulation of mature T lymphocytes (i.e., MHC class II-restricted T cells, including most T helper cells), and a subset of NK-T cells. In addition, CD4 has also been reported to be detectable on pluripotent hematopoietic stem cells, bone marrow myeloid and B-lymphocyte precursors, intrathymic lymphoid precursors, and a subset of splenic dendritic cells. CD4 has also been reported to be expressed on the plasma membrane of mouse egg cells and is involved in adhesion of the egg to MHC class II-bearing sperm.  CD4 is an antigen coreceptor on the T-cell surface which interacts with MHC class II molecules on antigen-presenting cells. It participates in T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase lck. The GK1.5 antibody reportedly blocks binding of the RM4-5 and H129.19, but not RM4-4 mouse CD4-specific antibodies.

      571146 Rev. 1
      Format Details
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      RY610
      The BD Horizon RealYellow™ 610 (RY610) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 610-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY610 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY610 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 Yellow-Green laser (561-nm), it can be used in conjunction with PE-CF594.
      altImg
      RY610
      557 nm
      610 nm
      571146 Rev.1
      Citations & References
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      View product citations for antibody "571146" on CiteAb

      Development References (10)

      1. Bosselut R, Zhang W, Ashe JM, Kopacz JL, Samelson LE, Singer A. Association of the adaptor molecule LAT with CD4 and CD8 coreceptors identifies a new coreceptor function in T cell receptor signal transduction. J Exp Med. 1999; 190(10):1517-1526. (Clone-specific: (Co)-stimulation, Immunoprecipitation). View Reference
      2. Dialynas DP, Quan ZS, Wall KA, et al. Characterization of the murine T cell surface molecule, designated L3T4, identified by monoclonal antibody GK1.5: similarity of L3T4 to the human Leu-3/T4 molecule. J Immunol. 1983; 131(5):2445-2451. (Immunogen: Blocking, Depletion, Flow cytometry). View Reference
      3. Dialynas DP, Wilde DB, Marrack P, et al. Characterization of the murine antigenic determinant, designated L3T4a, recognized by monoclonal antibody GK1.5: expression of L3T4a by functional T cell clones appears to correlate primarily with class II MHC antigen-reactivity. Immunol Rev. 1983; 74:29-56. (Clone-specific: Blocking, Depletion, Flow cytometry). View Reference
      4. Frederickson GG, Basch RS. L3T4 antigen expression by hemopoietic precursor cells. J Exp Med. 1989; 169(4):1473-1478. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
      5. Ghobrial RR, Boublik M, Winn HJ, Auchincloss H Jr. In vivo use of monoclonal antibodies against murine T cell antigens. Clin Immunol Immunopathol. 1989; 52(3):486-506. (Biology). View Reference
      6. Janeway CA Jr. The T cell receptor as a multicomponent signalling machine: CD4/CD8 coreceptors and CD45 in T cell activation. Annu Rev Immunol. 1992; 10:645-674. (Biology). View Reference
      7. Kruisbeek AM. In vivo depletion of CD4- and CD8-specific T cells. Curr Protoc Immunol. 1991; 4.1.1-4.1.5. (Clone-specific: Cytotoxicity, Depletion). View Reference
      8. Wineman JP, Gilmore GL, Gritzmacher C, Torbett BE, Muller-Sieburg CE. CD4 is expressed on murine pluripotent hematopoietic stem cells. Blood. 1992; 180(7):1717-1724. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting, Immunofluorescence, Immunoprecipitation). View Reference
      9. Wu L, Antica M, Johnson GR, Scollay R, Shortman K. Developmental potential of the earliest precursor cells from the adult mouse thymus. J Exp Med. 1991; 174(6):1617-1627. (Biology). View Reference
      10. Zheng B, Han S, Kelsoe G. T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1. J Exp Med. 1996; 184(3):1083-1091. (Clone-specific: Immunohistochemistry). View Reference
      View All (10) View Less
      571146 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.