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BD Horizon™ BV650 Mouse Anti-Human CD25
Clone M-A251 (RUO)
Flow cytometric analysis of CD25 expression on unstimulated and stimulated human peripheral blood lymphocytes.
Left Panel: Whole blood was stained with either BD Horizon™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram) or BD Horizon™ BV650 Mouse Anti-Human CD25 antibody (Cat. No. 563718/563719; 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™ BV650 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV650 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™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram) or BD Horizon™ BV650 Mouse Anti-Human CD25 antibody (Cat. No. 563718/563719; 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™ BV650 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV650 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™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram) or BD Horizon™ BV650 Mouse Anti-Human CD25 antibody (Cat. No. 563718/563719; 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™ BV650 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV650 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™ BV650 Mouse IgG1, κ Isotype Control (Cat. No. 563231; dashed line histogram) or BD Horizon™ BV650 Mouse Anti-Human CD25 antibody (Cat. No. 563718/563719; 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™ BV650 Mouse Anti-Human CD25 antibody (solid line histogram) or with BD Horizon™ BV650 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
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
AB_2744337
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™ BV650 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV650 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
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- BD Horizon Brilliant Violet 650 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
BV650 Mouse IgG1, k Isotype Control RUO
Size
50 µg
Cat No.
563231
BV650 Mouse Anti-Human CD25 RUO
Size
25 Tests
Cat No.
563718
563719 Rev. 2
Antibody Details
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™ BV650 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 650-nm. BD Horizon™ BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
563719 Rev. 2
Format Details
BV650
The BD Horizon Brilliant Violet™ 650 (BV650) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 406-nm and an acceptor dye with an emission maximum (Em Max) at 649-nm. BV650, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 650-nm (e.g., a 660/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) 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.
BV650
406 nm
649 nm
563719 Rev.2
Citations & References
View product citations for antibody "563719" on CiteAb
Development References (6)
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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.
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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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
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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.
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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.
-
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.
563719 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.