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BV421 Mouse Anti-Human CD25 2A3 RUO

BV421 Mouse Anti-Human CD25 

Clone  2A3   (RUO)

  • Brand BD Horizon™
  • Alternative Name IL-2R; IL2RA; IL-2Rα; TCGFR; TAC antigen; p55
  • Vol. Per Test 5 µl
  • Isotype Mouse BALB/c IgG1, κ
  • Reactivity Human (QC Testing) 
  • Application Flow cytometry (Routinely Tested) 
  • Immunogen Human Phytohemagglutinin-activated T Cells
  • Workshop No. III A769,T153; IV A8
  • Entrez Gene ID 3559 
  • Storage Buffer Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

The 2A3 monoclonal antibody specifically binds to human CD25, the low-affinity alpha subunit of the Interleukin-2 Receptor (IL-2Rα). CD25 associates with CD122 (IL-2R β chain) and CD132 (common γ chain or γc) to form the high-affinity signal-transducing IL-2R complex. CD25 is expressed by a subset of peripheral blood lymphocytes including CD4+CD25+ natural regulatory T cells. CD25 antigen density increases on activated T cells including phytohemagglutinin (PHA)-, concanavalin A (Con A)-, and CD3-activated T lymphocytes. High levels of CD25 can also be expressed by T lymphocytes from mixed lymphocyte cultures and by human T-lymphocyte leukemia virus (HTLV)-infected T-lymphocyte leukemia lines, for example, HUT-102. Recombinant IL-2 blocks the binding of the 2A3 antibody to PHA-activated T lymphocytes.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

Format
  • Format BV421
  • Excitation Source Violet 405 nm
  • Excitation Max 407 nm
  • Emission Max 421 nm

BV421 is a polymer-based dye excited by the violet laser and is one of the brightest fluorochromes offered by BD Biosciences. Conjugates are typically 10 times brighter than Pacific Blue™ conjugates and are often as bright as or brighter than PE conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously.

Other Formats →

Suggested Companion Products

Lysing Solution 10X Concentrate CE/IVD
100 Cat No: 349202

Lysing Buffer RUO
100 mL Cat No: 555899

Stain Buffer (FBS) RUO
500 mL Cat No: 554656

Stain Buffer (BSA) RUO
500 mL Cat No: 554657

BV421 Mouse IgG1, k Isotype Control RUO
50 µg Cat No: 562438

Resources & Tools
 Spectrum Viewer  Panel Designer  Spectrum Viewer  Download TDS  Regulatory Document Website
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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. Brilliant Violet™ 421 is a trademark of Sirigen.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.


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  2. Greene WC, Leonard WJ. The human interleukin-2 receptor. Annu Rev Immunol. 1986; 4:69-95. View reference

  3. Lando Z, Sarin P, Megson M, et al. Association of human T-cell leukaemia/lymphoma virus with the Tac antigen marker for the human T-cell growth factor receptor. Nature. 1983; 305(5936):733-736. View reference

  4. Leonard WJ, Depper JM, Uchiyama T, Smith KA, Waldmann TA, Greene WC. A monoclonal antibody that appears to recognize the receptor for human T-cell growth factor; partial characterization of the receptor. Nature. 1982; 300(5889):267-269. View reference

  5. Ng WF, Duggan PJ, Ponchel F, et al. Human CD4(+)CD25(+) cells: a naturally occurring population of regulatory T cells. Blood. 2001; 98(9):2736-2744. View reference

  6. Rambaldi A, Young DC, Herrmann F, Cannistra SA, Griffin JD. Interferon-gamma induces expression of the interleukin 2 receptor gene in human monocytes. Eur J Immunol. 1987; 17(1):153-156. View reference

  7. Robb RJ, Greene WC, Rusk CM. Low and high affinity cellular receptors for interleukin 2. Implications for the level of Tac antigen. J Exp Med. 1984; 160(4):1126-1146. View reference

  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.

  9. Sereti I, Martinez-Wilson H, Metcalf JA, et al. Long-term effects of intermittent interleukin 2 therapy in patients with HIV infection: characterization of a novel subset of CD4(+)/CD25(+) T cells. Blood. 2002; 100(6):2159-2167. View reference

  10. Siegel JP, Sharon M, Smith PL, Leonard WJ. The IL-2 receptor beta chain (p70): role in mediating signals for LAK, NK, and proliferative activities. Science. 1987; 238(4823):75-78. View reference

  11. Teshigawara K, Wang HM, Kato K, Smith KA. Interleukin 2 high-affinity receptor expression requires two distinct binding proteins. J Exp Med. 1987; 165(1):223-238. View reference

  12. Urdal DL, March CJ, Gillis S, Larsen A, Dower SK. Purification and chemical characterization of the receptor for interleukin 2 from activated human T lymphocytes and from a human T-cell lymphoma cell line. Proc Natl Acad Sci U S A. 1984; 81(20):6481-6485. View reference

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