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      BUV805 Rat Anti-Mouse CD25 (IL-2 Receptor α)
      BUV805 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.        Left and Middle Plots: Mouse splenic leukocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047) and with either no antibody (Autofluorescence Control; Left Plot) or BD Horizon™ BUV805 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (Cat. No. 569603/569604; Middle Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) versus CD4 was derived from 7-AAD negative-gated events with the forward and side light-scatter characteristics of viable leukocytes.          Right Plot: Mouse splenic leukocytes were stimulated for 3 days with Concanavalin A (ConA). The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody and then stained with BD Horizon BUV805 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (solid line histogram) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The fluorescence histogram showing CD25 (IL-2 Receptor α) expression (or Autofluorescence) was derived from 7-AAD negative-events with the forward and side light-scatter characteristics of viable lymphoblasts.        Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      BUV805 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.        Left and Middle Plots: Mouse splenic leukocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047) and with either no antibody (Autofluorescence Control; Left Plot) or BD Horizon™ BUV805 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (Cat. No. 569603/569604; Middle Plot) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) versus CD4 was derived from 7-AAD negative-gated events with the forward and side light-scatter characteristics of viable leukocytes.          Right Plot: Mouse splenic leukocytes were stimulated for 3 days with Concanavalin A (ConA). The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody and then stained with BD Horizon BUV805 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (solid line histogram) at 0.5 µg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The fluorescence histogram showing CD25 (IL-2 Receptor α) expression (or Autofluorescence) was derived from 7-AAD negative-events with the forward and side light-scatter characteristics of viable lymphoblasts.        Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      製品詳細
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      BD Horizon™
      Interleukin-2 receptor alpha chain; IL-2RA; IL-2Rα; Il2ra; IL-2R p55
      Mouse (QC Testing)
      Rat OFA, also known as Outbred OFA IgG1, λ
      IL-2-dependent cytolytic mouse T-cell clone B6.1
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      16184
      Aqueous buffered solution containing ≤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 unconjugated antibody and free dye were removed.
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      推奨アッセイ手順

         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. This will 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).

         Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

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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. 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. 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 Ultraviolet 805 is covered by one or more of the following US patents: 8,110,673, 8,158,444; 8,227,187; 8,575,303; 8,354,239.
      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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      569603 Rev. 1
      抗体の詳細
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      PC61

      The PC61 monoclonal antibody specifically binds to CD25, the low-affinity IL-2 Receptor α chain (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 and 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 on the CD45R/B220low TdT- sIg- Pre-B/Pre-B-II and CD45R/B220low TdT- sIgM+ sIgD- immature B stages, but not on the CD45R/B220low TdT+ sIg- Pro-B/Pre-B-I stage nor on CD45R/B220high TdT- sIgM+ 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+ 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. The PC61 antibody recognizes an epitope of CD25 which is distinct from the IL-2 binding site and from those recognized by mAbs 3C7 and 7D4. It blocks binding of IL-2 to CD25, presumably by inducing a conformational change in CD25.

      569603 Rev. 1
      フォーマットの詳細
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      BUV805
      The BD Horizon Brilliant™ Ultraviolet 805 (BUV805) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 351-nm and an acceptor dye with an emission maximum (Em Max) at 803-nm. BUV805, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 805-nm (e.g., a 820/60 or a 780/60 bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BUV805
      Ultraviolet 355 nm
      351 nm
      803 nm
      569603 Rev.1
      引用＆参考文献
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      Development References (9)

      1. Ceredig R, Lowenthal JW, Nabholz M, MacDonald HR. Expression of interleukin-2 receptors as a differentiation marker on intrathymic stem cells. Nature. 1985; 314(6006):98-100. (Clone-specific: Blocking, Immunohistochemistry). View Reference
      2. 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. (Biology). View Reference
      3. Ernst DN, Weigle WO, McQuitty DN, Rothermel AL, Hobbs MV. Stimulation of murine T cell subsets with anti-CD3 antibody. Age-related defects in the expression of early activation molecules. J Immunol. 1989; 142(5):1413-1421. (Clone-specific: Flow cytometry). View Reference
      4. 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
      5. Godfrey DI, Zlotnik A. Control points in early T-cell development. Immunol Today. 1993; 14(11):547-553. (Biology). View Reference
      6. Lowenthal JW, Corthésy P, Tougne C, Lees R, MacDonald HR, Nabholz M. High and low affinity IL 2 receptors: analysis by IL 2 dissociation rate and reactivity with monoclonal anti-receptor antibody PC61. J Immunol. 1985; 135(6):3988-3994. (Immunogen: Bioassay, Blocking, Functional assay, Inhibition, Radioimmunoassay). View Reference
      7. Lowenthal JW, Zubler RH, Nabholz M, MacDonald HR. Similarities between interleukin-2 receptor number and affinity on activated B and T lymphocytes. Nature. 1985; 315(6021):669-672. (Clone-specific: Blocking, Immunoprecipitation, Radioimmunoassay). View Reference
      8. 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: Blocking). View Reference
      9. 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
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      569603 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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