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BV786 Mouse Anti-Human CD25
BV786 Mouse Anti-Human CD25
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.        Left Panel: Whole blood was stained with either BD Horizon™ BV786 Mouse IgG1, κ Isotype Control (Cat. No. 563330; dashed line histogram) or BD Horizon™ BV786 Mouse Anti-Human CD25 antibody (Cat. No. 563700/563701; solid line histogram). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).        Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV786 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV786 Mouse IgG1, κ Isotype Control (dashed 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 Panel) or lymphoblasts (Right Panel). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.        Left Panel: Whole blood was stained with either BD Horizon™ BV786 Mouse IgG1, κ Isotype Control (Cat. No. 563330; dashed line histogram) or BD Horizon™ BV786 Mouse Anti-Human CD25 antibody (Cat. No. 563700/563701; solid line histogram). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899).        Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV786 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV786 Mouse IgG1, κ Isotype Control (dashed 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 Panel) or lymphoblasts (Right Panel). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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
AB_2744338
Aqueous buffered solution containing BSA 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™ BV786 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV786 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD Optibuild Brilliant reagents) 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).

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Cy is a trademark of GE Healthcare.
  8. 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.
  9. BD Horizon Brilliant Violet 786 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563700 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™ BV786 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. This dye is a tandem fluorochrome of BD Horizon™ BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 786-nm.  BD Horizon™ BV786 can be excited by the violet laser and detected in a filter used to detect Cy7™-like dyes (eg, 780/60-nm filter).

563700 Rev. 1
Format Details
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BV786
The BD Horizon Brilliant Violet™ 786 (BV786) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an Ex Max of 407-nm and an acceptor dye with an Em Max at 786-nm.  BV786, driven by BD innovation, is designed to be excited by the violet laser and detected using a filter, centered near 785 nm (e.g. 780/60 nm bandpass filter).  Please ensure that your instrument’s configurations (lasers and filters) are appropriate for this dye.
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BV786
Violet 405 nm
407 nm
786 nm
563700 Rev.1
Citations & References
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View product citations for antibody "563700" on CiteAb

Development References (6)

  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. 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
  4. 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.
  5. 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.
  6. 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 (6) View Less
563700 Rev. 1

 

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