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      BV480 Rat Anti-Mouse CD25 (IL-2 Receptor α)
      BV480 Rat Anti-Mouse CD25 (IL-2 Receptor α)
      Flow cytometric analysis of CD25 (IL-2 Receptor α) expression on Unstimulated and Stimulated Mouse splenic leukocytes.    Unstimulated Cells (Left and Middle Plots): Freshly prepared Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The Unstimulated cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and with either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (Cat. No. 565649; Left Plot) or BD Horizon™ BV480 Rat Anti-Mouse CD25  (IL-2 Receptor α) antibody (Cat. No. 569544/569545; Middle Plot) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or Ig Isotype control staining] versus CD4 was generated for gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes.    Stimulated Cells (Right Plot): Mouse splenic leukocytes were stimulated with Concanavalin A (Con A) for 3 days. The Stimulated cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon™ BV480 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (solid line histogram) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphoblasts.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      BV480 Rat Anti-Mouse CD25 (IL-2 Receptor α)
      Flow cytometric analysis of CD25 (IL-2 Receptor α) expression on Unstimulated and Stimulated Mouse splenic leukocytes.    Unstimulated Cells (Left and Middle Plots): Freshly prepared Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The Unstimulated cells were then stained with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and with either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (Cat. No. 565649; Left Plot) or BD Horizon™ BV480 Rat Anti-Mouse CD25  (IL-2 Receptor α) antibody (Cat. No. 569544/569545; Middle Plot) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or Ig Isotype control staining] versus CD4 was generated for gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphocytes.    Stimulated Cells (Right Plot): Mouse splenic leukocytes were stimulated with Concanavalin A (Con A) for 3 days. The Stimulated cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (dashed line histogram) or BD Horizon™ BV480 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (solid line histogram) at 1 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) lymphoblasts.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      제품 세부 정보
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      BD Horizon™
      Ms CD25 BV480 3C7 100ug
      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
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      준비 및 보관

      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 unconjugated antibody and free dye were removed.
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      권장 분석 절차

         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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      제품 고시

      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. 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. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. 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.
      8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      9. For U.S. patents that may apply, see bd.com/patents.
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      569544 Rev. 1
      항체 세부 정보
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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.

      569544 Rev. 1
      형광 세부 정보
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      BV480
      The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV480
      Violet 405 nm
      440 nm
      479 nm
      569544 Rev.1
      인용 및 참고 문헌
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      개발 참고 자료 (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). 참조 보기
      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). 참조 보기
      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). 참조 보기
      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. (Clone-specific: Flow cytometry). 참조 보기
      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). 참조 보기
      6. Malek TR. The biology of interleukin-2. Annu Rev Immunol. 2008; 26:453-479. (Biology). 참조 보기
      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). 참조 보기
      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). 참조 보기
      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). 참조 보기
      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). 참조 보기
      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). 참조 보기
      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). 참조 보기
      모두 보기 (12) 간단히 보기
      569544 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.

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