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Alexa Fluor™ 488 Mouse Anti-Human CD25
Alexa Fluor™ 488 Mouse Anti-Human CD25
Multiparameter flow cytometric analysis of CD25 expression on unstimulated Human leucocytes (Left Plots) and stimulated (Right Plot) Human lymphocytes.    Left Plots- Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425) and with either Alexa Fluor™488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD25 antibody (Cat. No. 568871/568872; Right Plot). 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 intact lymphocytes.    Right Plot - Peripheral blood mononuclear cells (PBMC) were stimulated for 3 days with Phytohemagglutinin (PHA) and then stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-Human CD25 antibody (Cat. No. 568871/568872; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescent histogram showing CD25 expression (or Ig Isotype control staining) was derived from gated events with light-scatter characteristics of viable (DAPI-negative) lymphoblasts.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CD25 expression on unstimulated Human leucocytes (Left Plots) and stimulated (Right Plot) Human lymphocytes.    Left Plots- Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD4 antibody (Cat. No. 562424/562425) and with either Alexa Fluor™488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD25 antibody (Cat. No. 568871/568872; Right Plot). 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 intact lymphocytes.    Right Plot - Peripheral blood mononuclear cells (PBMC) were stimulated for 3 days with Phytohemagglutinin (PHA) and then stained with either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; dashed line histogram) or Alexa Fluor™ 488 Mouse Anti-Human CD25 antibody (Cat. No. 568871/568872; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescent histogram showing CD25 expression (or Ig Isotype control staining) was derived from gated events with light-scatter characteristics of viable (DAPI-negative) lymphoblasts.    Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
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BD Pharmingen™
IL-2Rα; IL2RA; TAC antigen; TCGFR; p55
Human (QC Testing)
Mouse BALB/c IgG1, κ
Recombinant Human CD25
Flow cytometry (Routinely Tested)
5 µl
V C012
3559
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 to the dye under optimum conditions and unreacted dye was 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 to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
568871 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.

568871 Rev. 1
フォーマットの詳細
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
568871 Rev.1
引用&参考文献
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Development References (6)

  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, Gel shift). View Reference
  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. Li H, Zhou R, Wang C, et al. T follicular regulatory cells infiltrate the human airways during the onset of acute respiratory distress syndrome and regulate the development of B regulatory cells.. Immunol Res. 2018; 66(4):548-554. (Clone-specific: Flow cytometry). View Reference
  4. Nie H, Zheng Y, Li R, Zhang J. Reply to Suppressive activity of human regulatory T cells is maintained in the presence of TNF.. Nat Med. 2016; 22(1):18-9. (Clone-specific: Flow cytometry). View Reference
  5. 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). View Reference
  6. 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 (6) 表示項目を減らす
568871 Rev. 1

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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.