BV605 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/test
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing)
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
Regulatory Status Legend
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-2Rβ chain) and CD132 (common γ chain or γc) to form the high-affinity signal-transducing IL-2R complex. CD25 is expressed by subsets of thymocytes and peripheral blood lymphocytes including CD4+CD25+ regulatory T cells and memory 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 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. CD25 can also be expressed by activated B cells and macrophages. Recombinant IL-2 blocks the binding of the 2A3 antibody to PHA-activated T lymphocytes.
This antibody is conjugated to BD Horizon BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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).
BV605 is a tandem fluorochrome that combines BD Horizon BV421 and an acceptor dye with an Em max at 602-nm. These conjugates are very bright, often exhibiting similar brightness to equivalent PE conjugates. Due to the excitation of acceptor dye by the green (532-nm) and yellow-green (561-nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers.
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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™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.
- 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.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Ficoll-Paque is a trademark of Amersham Biosciences Limited.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- CF™ is a trademark of Biotium, Inc.