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BUV737 Mouse Anti-Human CD25
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This product is the replacement for [564386].
BUV737 Mouse Anti-Human CD25
Flow cytometric analysis of CD25 expression on unstimulated peripheral blood lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD4 antibody (555347/561843/561844) and either BD Horizon™ BUV737 Mouse IgG1, κ Isotype Control (Cat. No. 612758; Left Plot) or BD Horizon BUV737 Mouse Anti-Human CD25 antibody (Cat. No. 612806/612807; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric contour plot showing the correlated expression of CD25 (or Ig Isotype control staining) versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
BUV737 Mouse Anti-Human CD25
Flow cytometric analysis of CD25 expression on activated peripheral blood lymphocytes.  Human peripheral blood mononuclear cells were stimulated for 3 days with Phytohemagglutinin. The cells were stained with either BD Horizon BUV737 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon BUV737 Mouse Anti-Human CD25 antibody (solid line histogram). The fluorescence histogram showing CD25 expression (or Ig Isotype control staining) was derived from gated 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.
Flow cytometric analysis of CD25 expression on unstimulated peripheral blood lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD4 antibody (555347/561843/561844) and either BD Horizon™ BUV737 Mouse IgG1, κ Isotype Control (Cat. No. 612758; Left Plot) or BD Horizon BUV737 Mouse Anti-Human CD25 antibody (Cat. No. 612806/612807; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric contour plot showing the correlated expression of CD25 (or Ig Isotype control staining) versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD25 expression on activated peripheral blood lymphocytes.  Human peripheral blood mononuclear cells were stimulated for 3 days with Phytohemagglutinin. The cells were stained with either BD Horizon BUV737 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon BUV737 Mouse Anti-Human CD25 antibody (solid line histogram). The fluorescence histogram showing CD25 expression (or Ig Isotype control staining) was derived from gated 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.
Product Details
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BD Horizon™
IL-2R; IL2RA; IL-2Rα; TCGFR; TAC antigen; p55
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Phytohemagglutinin-activated T Cells
Flow cytometry (Routinely Tested)
5 µl
III A769,T153; IV A8
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 with BD Horizon BUV737 under optimum conditions, and unconjugated antibody and free BD Horizon BUV737 were 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 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 two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  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).

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. BD Horizon Brilliant Ultraviolet 737 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.
  6. 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.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
612807 Rev. 2
Antibody Details
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2A3

The 2A3 monoclonal antibody specifically binds to human CD25, the low-affinity alpha subunit of the Interleukin-2 Receptor (IL- 2Rα). CD25 associates with CD122 (IL-2Rβ chain) and CD132 (common γ chain or γc) to form the high-affinity signal-transducing IL-2R complex. CD25 is expressed by subsets of thymocytes and peripheral blood lymphocytes including CD4+CD25+ regulatory T cells and memory T cells. CD25 antigen density increases on activated T cells including phytohemagglutinin (PHA)-, concanavalin A (Con A)-, and CD3-activated T lymphocytes. High levels of CD25 can be expressed by T lymphocytes from mixed lymphocyte cultures and by human T-lymphocyte leukemia virus (HTLV)-infected T-lymphocyte leukemia lines, for example, HUT-102. CD25 can also be expressed by activated B cells and macrophages. Recombinant IL-2 blocks the binding of the 2A3 antibody to PHA-activated T lymphocytes.

The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 737 nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 nm filter. Due to the excitation of the acceptor dye by the red laser line, there may be significant spillover into red laser detectors with filters in the 700-720 nm range.

612807 Rev. 2
Format Details
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BUV737
The BD Horizon Brilliant™ Ultraviolet 737 (BUV737) 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 350-nm and an acceptor dye with an emission maximum (Em Max) at 735-nm. BUV737, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 740-nm (e.g., 740/35 bandpass filter). The acceptor dye can be excited by the Red (628–640nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BUV737
Ultraviolet 355 nm
350 nm
735 nm
612807 Rev.2
Citations & References
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Development References (13)

  1. Dower SK, Hefeneider SH, Alpert AR, Urdal DL. Quantitative measurement of human interleukin 2 receptor levels with intact and detergent-solubilized human T-cells. Mol Immunol. 1985; 22(8):937-947. (Clone-specific). View Reference
  2. Greene WC, Leonard WJ. The human interleukin-2 receptor. Annu Rev Immunol. 1986; 4:69-95. (Clone-specific). View Reference
  3. Jackson AL, Matsumoto H, Janszen M, Maino V, Blidy A, Shye S. Restricted expression of p55 interleukin 2 receptor (CD25) on normal T cells. Clin Immunol Immunopathol. 1990; 54(1):126-133. (Clone-specific: Flow cytometry). View Reference
  4. Lando Z, Sarin P, Megson M, et al. Association of human T-cell leukaemia/lymphoma virus with the Tac antigen marker for the human T-cell growth factor receptor. Nature. 1983; 305(5936):733-736. (Biology). View Reference
  5. Leonard WJ, Depper JM, Uchiyama T, Smith KA, Waldmann TA, Greene WC. A monoclonal antibody that appears to recognize the receptor for human T-cell growth factor; partial characterization of the receptor. Nature. 1982; 300(5889):267-269. (Biology). View Reference
  6. Ng WF, Duggan PJ, Ponchel F, et al. Human CD4(+)CD25(+) cells: a naturally occurring population of regulatory T cells. Blood. 2001; 98(9):2736-2744. (Biology). View Reference
  7. Rambaldi A, Young DC, Herrmann F, Cannistra SA, Griffin JD. Interferon-gamma induces expression of the interleukin 2 receptor gene in human monocytes. Eur J Immunol. 1987; 17(1):153-156. (Clone-specific). View Reference
  8. Robb RJ, Greene WC, Rusk CM. Low and high affinity cellular receptors for interleukin 2. Implications for the level of Tac antigen. J Exp Med. 1984; 160(4):1126-1146. (Biology). View Reference
  9. 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.
  10. Sereti I, Martinez-Wilson H, Metcalf JA, et al. Long-term effects of intermittent interleukin 2 therapy in patients with HIV infection: characterization of a novel subset of CD4(+)/CD25(+) T cells. Blood. 2002; 100(6):2159-2167. (Clone-specific: Flow cytometry). View Reference
  11. Siegel JP, Sharon M, Smith PL, Leonard WJ. The IL-2 receptor beta chain (p70): role in mediating signals for LAK, NK, and proliferative activities. Science. 1987; 238(4823):75-78. (Biology). View Reference
  12. Teshigawara K, Wang HM, Kato K, Smith KA. Interleukin 2 high-affinity receptor expression requires two distinct binding proteins. J Exp Med. 1987; 165(1):223-238. (Biology). View Reference
  13. Urdal DL, March CJ, Gillis S, Larsen A, Dower SK. Purification and chemical characterization of the receptor for interleukin 2 from activated human T lymphocytes and from a human T-cell lymphoma cell line. Proc Natl Acad Sci U S A. 1984; 81(20):6481-6485. (Immunogen: Blocking, Dot Blot, Immunoaffinity chromatography, Inhibition, Radioimmunoassay). View Reference
View All (13) View Less
612807 Rev. 2

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