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BV510 Mouse Anti-Human CD25
BV510 Mouse Anti-Human CD25
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.        Left and Middle Panels: Whole blood was stained with either BD Horizon™ BV510 Mouse IgG1, κ Isotype Control (Cat. No. 562946; Left Panel) or BD Horizon™ BV510 Mouse Anti-Human CD25 antibody (Cat. No. 563352/563351; Middle Panel). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two-color flow cytometric dot plot shows the correlated expression of CD25 (or Ig Isotype control staining) versus cellular autofluorescence derived from gated events with the forward light-scatter characteristics of viable lymphocytes.        Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV510 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV510 Mouse IgG1, κ Isotype Control (dashed 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™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.        Left and Middle Panels: Whole blood was stained with either BD Horizon™ BV510 Mouse IgG1, κ Isotype Control (Cat. No. 562946; Left Panel) or BD Horizon™ BV510 Mouse Anti-Human CD25 antibody (Cat. No. 563352/563351; Middle Panel). The erythrocytes were subsequently lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two-color flow cytometric dot plot shows the correlated expression of CD25 (or Ig Isotype control staining) versus cellular autofluorescence derived from gated events with the forward light-scatter characteristics of viable lymphocytes.        Right Panel: Phytohemagglutinin-stimulated (3 days) human peripheral blood mononuclear cells were stained with either BD Horizon™ BV510 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV510 Mouse IgG1, κ Isotype Control (dashed 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™ 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_2744336
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™ BV510 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV510 were removed.

Recommended Assay Procedures

Use of Anti-CD25 antibodies conjugated with brighter fluorochromes, eg, phycoerythrin, allophycocyanin, PE-Cy™7, or BD Horizon™ BV421, are recommended for staining cells that express relatively low levels of CD25 such as unstimulated regulatory T cells or memory T cells.

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. 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.
  8. BD Horizon Brilliant Violet 510 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563352 Rev. 2
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™ BV510 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 405-nm and Em Max at 510-nm, BD Horizon™ BV510 can be excited by the violet laser and detected in the BD Horizon™ V500 (525/50-nm) filter set. BD Horizon™ BV510 conjugates are useful for the detection of dim markers off the violet laser.

563352 Rev. 2
Format Details
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BV510
The BD Horizon Brilliant Violet™ 510 (BV510) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye with an excitation maximum (Ex Max) at 327-nm / 405-nm and an emission maximum (Em Max) at 512-nm. BV510, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 510-nm (e.g., a 525/50 bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV510
Violet 405 nm
327 nm, 405 nm
512 nm
563352 Rev.2
Citations & References
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Development References (3)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  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. 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.
563352 Rev. 2

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.