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      BV786 Rat Anti-Mouse CD25
      BV786 Rat Anti-Mouse CD25
      Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes. Left and Middle Panels: Freshly prepared mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051/561091) and either BD Horizon™ BV786 Rat IgG2b, κ Isotype Control (Cat. No. 563334; Left Panel) or BD Horizon BV786 Rat Anti-Mouse CD25 antibody (Cat. No. 564368; Middle Panel). Two-color flow cytometric contour plots showing the correlated expression patterns for CD25 (or Ig Isotype Control staining) versus CD4 were generated for gated events with the forward and side light- scatter characteristics of viable leucocytes. Right Panel: Mouse splenic leucocytes were stimulated with concanavalin A for 3 days. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon BV786 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon BV786 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      BV786 Rat Anti-Mouse CD25
      Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes. Left and Middle Panels: Freshly prepared mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051/561091) and either BD Horizon™ BV786 Rat IgG2b, κ Isotype Control (Cat. No. 563334; Left Panel) or BD Horizon BV786 Rat Anti-Mouse CD25 antibody (Cat. No. 564368; Middle Panel). Two-color flow cytometric contour plots showing the correlated expression patterns for CD25 (or Ig Isotype Control staining) versus CD4 were generated for gated events with the forward and side light- scatter characteristics of viable leucocytes. Right Panel: Mouse splenic leucocytes were stimulated with concanavalin A for 3 days. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon BV786 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon BV786 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Horizon™
      Interleukin-2 receptor alpha chain; IL-2RA; IL-2Rα; Il2ra; IL-2R p55
      Mouse (QC Testing)
      Rat LEW, also known as Lewis IgG2b, κ
      IL-2-dependent BALB/c mouse cell line
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      16184
      AB_2738771
      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 BD Horizon™ BV786 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV786 were removed.
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      Recommended Assay Procedures

      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
      7. Cy is a trademark of GE Healthcare.
      8. BD Horizon Brilliant Violet 786 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564368 Rev. 2
      Antibody Details
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      3C7

      The 3C7 monoclonal antibody specifically binds to CD25, the low affinity IL-2 Receptor (IL-2Rα, p55) expressed on activated T and B lymphocytes from all mouse strains tested. IL-2Rα by itself is not a signaling receptor. However, it can combine with IL-2 Receptor β (CD122) and γc (CD132) chains to form high-affinity signaling receptor complexes for IL-2. Resting T and B lymphocytes as well as resting and activated NK cells do not express IL-2Rα. CD25 is transiently expressed at a low level during normal B-cell development in the bone marrow during the CD45R/B220low TdT- sIg- Pre-B/Pre-B-II and CD45R/B220low TdT- sIgM+ sIgD- immature B stages, but not during the CD45R/B220low TdT+ sIg- Pro-B/Pre B-I stage nor on CD45R/B220high TdTsIgM+ sIgD+ mature B cells. It is expressed at a higher level during a very early stage of T-cell development in fetal and adult thymus. Peripheral CD25+ CD4+ T lymphocytes called regulatory T (Treg) cells are involved in the maintenance of self-tolerance. It has also been reported that dendritic cells express CD25 (recognized by mAb 7D4, another CD25-specific antibody). The 3C7 antibody recognizes an epitope of CD25 which is distinct from those recognized by the other CD25-specific mAbs, 7D4 and PC61. 3C7 blocks the binding of IL-2 to CD25.

      564368 Rev. 2
      Format Details
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      BV786
      The BD Horizon Brilliant Violet™ 786 (BV786) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an Ex Max of 407-nm and an acceptor dye with an Em Max at 786-nm.  BV786, driven by BD innovation, is designed to be excited by the violet laser and detected using a filter, centered near 785 nm (e.g. 780/60 nm bandpass filter).  Please ensure that your instrument’s configurations (lasers and filters) are appropriate for this dye.
      altImg
      BV786
      Violet 405 nm
      407 nm
      786 nm
      564368 Rev.2
      Citations & References
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      Development References (12)

      1. Chen J, Ma A, Young F, Alt FW. IL-2 receptor alpha chain expression during early B lymphocyte differentiation. Int Immunol. 1994; 6(8):1265-1268. (Clone-specific: Flow cytometry). View Reference
      2. Crowley M, Inaba K, Witmer-Pack M, Steinman RM. The cell surface of mouse dendritic cells: FACS analyses of dendritic cells from different tissues including thymus. Cell Immunol. 1989; 118(1):108-125. (Biology). View Reference
      3. Garni-Wagner BA, Witte PL, Tutt MM, et al. Natural killer cells in the thymus. Studies in mice with severe combined immune deficiency. J Immunol. 1990; 144(3):796-803. (Biology). View Reference
      4. Habu S, Okumura K, Diamantstein T, Shevach EM. Expression of interleukin 2 receptor on murine fetal thymocytes. Eur J Immunol. 1985; 15(5):456-460. (Biology). View Reference
      5. Malek TR, Robb RJ, Shevach EM. Identification and initial characterization of a rat monoclonal antibody reactive with the murine interleukin 2 receptor-ligand complex. Proc Natl Acad Sci U S A. 1983; 80(18):5694-5698. (Biology). View Reference
      6. Malek TR. The biology of interleukin-2. Annu Rev Immunol. 2008; 26:453-479. (Biology). View Reference
      7. Moreau JL, Nabholz M, Diamantstein T, Malek T, Shevach E, Theze J. Monoclonal antibodies identify three epitope clusters on the mouse p55 subunit of the interleukin 2 receptor: relationship to the interleukin 2-binding site. Eur J Immunol. 1987; 17(7):929-935. (Clone-specific: Bioassay, Blocking, Functional assay, Inhibition, Neutralization, Radioimmunoassay). View Reference
      8. Ortega G, Robb RJ, Shevach EM, Malek TR. The murine IL 2 receptor. I. Monoclonal antibodies that define distinct functional epitopes on activated T cells and react with activated B cells. J Immunol. 1984; 133(4):1970-1975. (Immunogen: Blocking, Flow cytometry, Immunoprecipitation, Inhibition, Neutralization, Radioimmunoassay). View Reference
      9. Pollard AM, Lipscomb MF. Characterization of murine lung dendritic cells: similarities to Langerhans cells and thymic dendritic cells. J Exp Med. 1990; 172(1):159-167. (Biology). View Reference
      10. Read S, Malmstrom V, Powrie F. Cytotoxic T lymphocyte-associated antigen 4 plays an essential role in the function of CD25(+)CD4(+) regulatory cells that control intestinal inflammation. J Exp Med. 2000; 192(2):295-302. (Biology). View Reference
      11. Rolink A, Grawunder U, Winkler TH, Karasuyama H, Melchers F. IL-2 receptor alpha chain (CD25, TAC) expression defines a crucial stage in pre-B cell development. Int Immunol. 1994; 6(8):1257-1264. (Biology). View Reference
      12. Takahashi T, Tagami T, Yamazaki S, et al. Immunologic self-tolerance maintained by CD25(+)CD4(+) regulatory T cells constitutively expressing cytotoxic T lymphocyte-associated antigen 4. J Exp Med. 2000; 192(2):303-309. (Biology). View Reference
      View All (12) View Less
      564368 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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