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      R718 Rat Anti-Mouse CD25 (IL-2 Receptor α)
      R718 Rat Anti-Mouse CD25 (IL-2 Receptor α)
      Multiparameter flow cytometric analysis of CD25 (IL-2 Receptor α) expression on unstimulated (Left Plots) and stimulated (Right Plot) Mouse splenic leukocytes.    Left and Middle Plots - 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/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553730) and with either no BD Horizon™ R718 conjugated antibody (Autofluorescence control; Left Plot ) or BD Horizon™ R718 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (Cat. No. 570355/570425; Middle Plot) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or cellular Autofluorescence) versus CD4 was derived from DAPI-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 either no BD Horizon™ R718 conjugated antibody (Autofluorescence control; dashed line histogram) or BD Horizon™ R718 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (solid line histogram) at 0.5 µg/test. DAPI Solution was added to cells right before analysis. The fluorescence histogram showing CD25 (IL-2 Receptor α) expression (or cellular Autofluorescence) was derived from DAPI-negative-gated events with the forward and side light-scatter characteristics of viable lymphoblasts.    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.
      R718 Rat Anti-Mouse CD25 (IL-2 Receptor α)
      Multiparameter flow cytometric analysis of CD25 (IL-2 Receptor α) expression on unstimulated (Left Plots) and stimulated (Right Plot) Mouse splenic leukocytes.    Left and Middle Plots - 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/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553730) and with either no BD Horizon™ R718 conjugated antibody (Autofluorescence control; Left Plot ) or BD Horizon™ R718 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (Cat. No. 570355/570425; Middle Plot) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or cellular Autofluorescence) versus CD4 was derived from DAPI-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 either no BD Horizon™ R718 conjugated antibody (Autofluorescence control; dashed line histogram) or BD Horizon™ R718 Rat Anti-Mouse CD25 (IL-2 Receptor α) antibody (solid line histogram) at 0.5 µg/test. DAPI Solution was added to cells right before analysis. The fluorescence histogram showing CD25 (IL-2 Receptor α) expression (or cellular Autofluorescence) was derived from DAPI-negative-gated events with the forward and side light-scatter characteristics of viable lymphoblasts.    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™
      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

      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. 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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
      8. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      9. For U.S. patents that may apply, see bd.com/patents.
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      570425 Rev. 1
      Antibody Details
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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.

      570425 Rev. 1
      Format Details
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      R718
      The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      R718
      Red 627-640 nm
      695 nm
      718 nm
      570425 Rev.1
      Citations & References
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      View product citations for antibody "570425" on CiteAb

      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
      View All (9) View Less
      570425 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.