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APC-R700 Rat Anti-Mouse CD25
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Consider BD Horizon™ Red 718 Reagents, bright small molecule alternative to APC-R700 with lower spread into the APC detector. More Info #
APC-R700 Rat Anti-Mouse CD25
Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes. Left and Middle Panels: Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/5 53142). The cells were then stained with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047/553046/561835) and either BD Horizon™ APC-R700 Rat IgG1, λ Isotype Control (Cat. No. 565133; Left Panel) or BD Horizon APC-R700 Rat Anti-Mouse CD25 antibody (Cat. No. 565134/565135; Middle Panel). Two-color flow cytometric dot plots showing the correlated expression of CD25 (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Right Panel: Mouse splenic leucocytes were stimulated for 3 days with concanavalin A. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody and then stained with either BD Horizon APC-R700 Rat IgG1, λ Isotype Control (dashed line histogram) or BD Horizon APC-R700 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated mouse splenocytes. Left and Middle Panels: Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/5 53142). The cells were then stained with FITC Rat Anti-Mouse CD4 antibody (Cat. No. 553047/553046/561835) and either BD Horizon™ APC-R700 Rat IgG1, λ Isotype Control (Cat. No. 565133; Left Panel) or BD Horizon APC-R700 Rat Anti-Mouse CD25 antibody (Cat. No. 565134/565135; Middle Panel). Two-color flow cytometric dot plots showing the correlated expression of CD25 (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of viable splenocytes. Right Panel: Mouse splenic leucocytes were stimulated for 3 days with concanavalin A. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody and then stained with either BD Horizon APC-R700 Rat IgG1, λ Isotype Control (dashed line histogram) or BD Horizon APC-R700 Rat Anti-Mouse CD25 antibody (solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer 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 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
AB_2744344
Aqueous buffered solution containing protein stabilizer 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 APC-R700 under optimum conditions, and unconjugated antibody and free BD Horizon APC-R700 were removed.

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. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  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 www.bdbiosciences.com/us/s/resources for technical protocols.
565134 Rev. 3
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.

This antibody was conjugated to BD Horizon APC-R700, which has been developed exclusively by BD Biosciences as a better alternative to Alexa Fluor® 700. APC-R700 excites and emits at similar wavelengths to Alexa Fluor® 700 yet exhibits significantly improved brightness. This dye can be excited by the red laser and detected with the same filter set as Alexa Fluor® (eg, 730/45-nm filter).

565134 Rev. 3
Format Details
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APC-R700
The BD Horizon™ APC-R700 (APC-R700) Dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of an Allophycocyanin (APC) dye donor that has excitation maximum (Ex Max) of 651-nm and an acceptor dye, R700, with an emission maximum (Em Max) at 706-nm. APC-R700, driven by BD innovation, is designed to be excited by the red (627–640-nm) laser and detected using an optical filter centered near 710-nm (e.g., a 720/40-nm bandpass filter). APC-R700 is a brighter alternative to Alexa Fluor™ 700. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC-R700
Red 627-640 nm
651 nm
706 nm
565134 Rev.3
Citations & References
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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, 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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565134 Rev. 3

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