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This product is the replacement for 740290.
569353Image1.png

Multiparameter flow cytometric analysis of CD25 (IL-2 Receptor α) expression on unstimulated Human leucocytes (Left Plots) and stimulated (Right Plot) Human lymphocytes.

   Left Plots- Human whole blood was stained with PE Mouse Anti-Human CD4 antibody (Cat. No. 561844/555347/561843) and with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; Right Plot) at 0.5 μg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or Ig Isotype control staining] versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

   Right Plot - Peripheral blood mononuclear cells (PBMC) were stimulated for 3 days with Phytohemagglutinin (PHA) and then stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescent histogram showing CD25 (IL-2 Receptor α) expression (or Ig Isotype control staining) was derived from gated events with light-scatter characteristics of viable (7-AAD-negative) lymphoblasts.

   Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

569353-2a.png

Multiparameter flow cytometric analysis of CD25 (IL-2 Receptor α) expression on unstimulated Human leucocytes (Left Plots) and stimulated (Right Plot) Human lymphocytes.

   Left Plots- Human whole blood was stained with PE Mouse Anti-Human CD4 antibody (Cat. No. 561844/555347/561843) and with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; Right Plot) at 0.5 μg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or Ig Isotype control staining] versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

   Right Plot - Peripheral blood mononuclear cells (PBMC) were stimulated for 3 days with Phytohemagglutinin (PHA) and then stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescent histogram showing CD25 (IL-2 Receptor α) expression (or Ig Isotype control staining) was derived from gated events with light-scatter characteristics of viable (7-AAD-negative) lymphoblasts.

   Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

569353Image1.png 569353-2a.png Violet-Cap_UV.png

Multiparameter flow cytometric analysis of CD25 (IL-2 Receptor α) expression on unstimulated Human leucocytes (Left Plots) and stimulated (Right Plot) Human lymphocytes.

   Left Plots- Human whole blood was stained with PE Mouse Anti-Human CD4 antibody (Cat. No. 561844/555347/561843) and with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; Right Plot) at 0.5 μg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or Ig Isotype control staining] versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

   Right Plot - Peripheral blood mononuclear cells (PBMC) were stimulated for 3 days with Phytohemagglutinin (PHA) and then stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescent histogram showing CD25 (IL-2 Receptor α) expression (or Ig Isotype control staining) was derived from gated events with light-scatter characteristics of viable (7-AAD-negative) lymphoblasts.

   Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Multiparameter flow cytometric analysis of CD25 (IL-2 Receptor α) expression on unstimulated Human leucocytes (Left Plots) and stimulated (Right Plot) Human lymphocytes.

   Left Plots- Human whole blood was stained with PE Mouse Anti-Human CD4 antibody (Cat. No. 561844/555347/561843) and with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; Right Plot) at 0.5 μg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD25 (IL-2 Receptor α) [or Ig Isotype control staining] versus CD4 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

   Right Plot - Peripheral blood mononuclear cells (PBMC) were stimulated for 3 days with Phytohemagglutinin (PHA) and then stained with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human CD25 (IL-2 Receptor α) antibody (Cat. No. 569353; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. The fluorescent histogram showing CD25 (IL-2 Receptor α) expression (or Ig Isotype control staining) was derived from gated events with light-scatter characteristics of viable (7-AAD-negative) lymphoblasts.

   Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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)
0.2 mg/ml
IV A053
3559
AB_3685000
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 unconjugated antibody and free dye were removed.
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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 to 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 BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).

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Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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 Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
  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. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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569353 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.

569353 Rev. 1
Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
348 nm
395 nm
569353 Rev.1
Citations & References
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View product citations for antibody "569353" on CiteAb

Development References (8)

  1. Herodin F, Thullier P, Garin D, Drouet M. Nonhuman primates are relevant models for research in hematology, immunology and virology. Eur Cytokine Netw. 2005; 16(2):104-116. (Clone-specific: Flow cytometry). View Reference
  2. 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.
  3. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  4. 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.
  5. 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
  6. 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.
  7. 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.
  8. 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 (8) View Less
569353 Rev. 1

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.