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R718 Mouse Anti-Human CD25
R718 Mouse Anti-Human CD25
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.        Left Plot: Whole blood was stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human CD25 antibody (Cat. No. 567065/567232; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).        Right Plot: Phytohemagglutinin (PHA)-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human CD25 antibody (solid line histogram).        The fluorescence histograms showing CD25 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes (Left Plot) or lymphoblasts (Right Plot). Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.        Left Plot: Whole blood was stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human CD25 antibody (Cat. No. 567065/567232; solid line histogram). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202).        Right Plot: Phytohemagglutinin (PHA)-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human CD25 antibody (solid line histogram).        The fluorescence histograms showing CD25 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes (Left Plot) or lymphoblasts (Right Plot). Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.
Product Details
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BD Horizon™
IL-2R; IL2RA; IL-2Rα; TCGFR; TAC antigen; p55
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Phytohemagglutinin stimulated human lymphocytes
Flow cytometry (Routinely Tested)
5 µl
IV A053
3559
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 unreacted dye was removed.

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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. 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.
  6. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567065 Rev. 1
Antibody Details
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M-A251

The M-A251 monoclonal antibody specifically binds to the 55 kDa type I transmembrane glycoprotein known as  low-affinity interleukin-2 receptor alpha chain subunit (IL-2Rα). CD25 is expressed on regulatory T cells,  activated lymphocytes (T and B), and monocytes. It associates with the IL-2Rβ/CD122 and IL-2Rγ/CD132 receptor chains to form the high-affinity IL-2R complex. CD25 expression on T and B lymphocytes is upregulated by antigenic or mitogenic stimulation. Soluble CD25/IL-2Rα is produced as a consequence of lymphocyte stimulation and is found in biological fluids following inflammatory responses.

The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567065 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.
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R718
Red 627-640 nm
695 nm
718 nm
567065 Rev.1
Citations & References
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View product citations for antibody "567065" on CiteAb

Development References (7)

  1. Janszen M, Buck D, Maino VC. Functional and molecular properties of CD25 monoclonal antibodies. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:403-406.
  2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  3. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
  4. Nakamura K, Kitani A, Fuss I, et al. TGF-β1 plays an important role in the mechanism of CD4+CD25+ regulatory T cell activity in both humans and mice. J Immunol. 2004; 172(2):834-842. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  5. Ravoet AM, Latinne D, Couvreur B, et al. CD25 mab: Epitopes recognized, effect on lymphocyte activation, mediation of ADCC. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:408-411.
  6. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
  7. Schwarting R, Stein H. Cluster report: CD25. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:399-403.
View All (7) View Less
567065 Rev. 1

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.