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BUV395 Mouse Anti-Human CD25
BUV395 Mouse Anti-Human CD25

Flow cytometric analysis of CD25 expression on unstimulated (Left Plots) and activated (Right Plot) human lymphocytes.

   Left Plots Whole blood was stained with APC Mouse Anti-Human CD4 antibody (Cat. No. 555349/561840/561841) and with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547) or BD Horizon™ BUV395 Mouse Anti-Human CD25 antibody (Cat. No. 567483/567484) as indicated. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density 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 viable lymphocytes.

   Right Plot Peripheral blood mononuclear cells 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 antibody (Cat. No. 567483/567484; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. 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 (7AAD-negative) lymphoblasts.

   Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Flow cytometric analysis of CD25 expression on unstimulated (Left Plots) and activated (Right Plot) human lymphocytes.

   Left Plots Whole blood was stained with APC Mouse Anti-Human CD4 antibody (Cat. No. 555349/561840/561841) and with either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547) or BD Horizon™ BUV395 Mouse Anti-Human CD25 antibody (Cat. No. 567483/567484) as indicated. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density 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 viable lymphocytes.

   Right Plot Peripheral blood mononuclear cells 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 antibody (Cat. No. 567483/567484; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. 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 (7AAD-negative) lymphoblasts.

   Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

製品詳細
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BD Horizon™
IL-2Rα; IL2RA; TAC antigen; TCGFR; p55
Human (QC Testing)
Mouse BALB/c IgG1, κ
Recombinant Human CD25
Flow cytometry (Routinely Tested)
5 µl
C012
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


調製と保管

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.

推奨アッセイ手順

     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 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).

製品通知

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567484 Rev. 1
抗体の詳細
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BC96

The BC96 monoclonal antibody specifically binds to human CD25, the low-affinity alpha subunit of the Interleukin-2 Receptor (IL-2Rα), which is also known as TAC antigen. IL2RA encodes CD25 which is a 55 kDa type I transmembrane glycoprotein comprised of an extracellular region with two Complement Control Protein domains (CCP) followed by a transmembrane region and a short cytoplasmic tail. CD25 is constitutively expressed at high levels on natural T regulatory cells and variably expressed on conventional T cells and B cells and their precursors, NK cells, monocytes, and macrophages. CD25 expression can be highly upregulated upon antigenic or mitogenic stimulation of T cells or B cells. A soluble form of CD25 is found in biological fluids due to proteolytic cleavage of the extracellular region of transmembrane CD25. CD25 noncovalently associates with CD122 (IL-2Rβ chain) and CD132 (IL-2Rγ, also known as the common γ chain or γc) to form the high-affinity signal-transducing IL-2R complex (IL-2Rαβγ).  This heterotrimeric receptor mediates biological activities of IL-2 which can act as a cellular activation, growth, and differentiation factor and regulator of cell viability. Analysis of CD25 expression can be used to characterize the nature of normal leucocytes in their resting states or activated during inflammatory or immune responses as well as those present in certain disease states.

The antibody was conjugated to BD Horizon BUV395 which has been exclusively developed by BD Biosciences as an optimal dye for use on a 355 nm laser equipped instrument. With an Ex Max at 348 nm  and an Em Max at 395 nm, this dye has virtually no spillover into any other detector. BD Horizon BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.

567484 Rev. 1
フォーマットの詳細
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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
Ultraviolet 355 nm
348 nm
395 nm
567484 Rev.1
引用&参考文献
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開発者向け参考資料 (4)

  1. Chapel A, Bensussan A, Vilmer E, Dormont D. Differential human immunodeficiency virus expression in CD4+ cloned lymphocytes: from viral latency to replication.. J Virol. 1992; 66(6):3966-70. (Immunogen: Flow cytometry). 参考文献を見る
  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. Poszepczynska E, Bagot M, Echchakir H, et al. Functional characterization of an IL-7-dependent CD4(+)CD8alphaalpha(+) Th3-type malignant cell line derived from a patient with a cutaneous T-cell lymphoma.. 2000; 96(3):1056-63. (Clone-specific). 参考文献を見る
  4. 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.
すべて表示する (4) 表示項目を減らす
567484 Rev. 1

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