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Two-color flow cytometric analysis of CD3 Molecular Complex expression on mouse splenocytes. Splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with PE Rat Anti-Mouse CD19 antibody (Cat. No. 557399/553786/561736) and either BD Horizon™ BV605 Rat IgG2b, κ Isotype Control (Cat. No. 563145; Left Panel) or BD Horizon™ BV605 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 564009; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of CD3 Molecular Complex (or Ig Isotype control staining) versus CD19 for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BV605 Rat Anti-Mouse CD3 Molecular Complex
Regulatory Statusの凡例
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation and Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- 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.
- CF™ is a trademark of Biotium, Inc.
- 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.
関連製品
The 17A2 monoclonal antibody specifically binds to the T-cell receptor-associated CD3 complex that is expressed on many thymocytes and mature T lymphocytes. Plate-bound 17A2 antibody has been reported to induce IL-2 production by cultured T cells in the absence of accessory cells. The binding of the 17A2 antibody to T cells can be blocked by the anti-CD3e mAb 145-2C11 (Cat. No. 557306/553058/550275). This suggests that the 17A2 antibody recognizes an epitope of the CD3 epsilon chain. In vivo treatment with 17A2 antibody has been reported to partially deplete T lymphocytes and temporarily down-modulates CD3 expression on T cells.
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).
Development References (3)
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Miescher GC, Schreyer M, MacDonald HR. Production and characterization of a rat monoclonal antibody against the murine CD3 molecular complex. Immunol Lett. 1989; 23(2):113-118. (Immunogen: Cytotoxicity, Depletion, Flow cytometry, Functional assay, Immunohistochemistry, Immunoprecipitation, Stimulation). View Reference
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Mysliwietz J, Thierfelder S. Antilymphocytic antibodies and marrow transplantation. XII. Suppression of graft-versus-host disease by T-cell-modulating and depleting antimouse CD3 antibody is most effective when preinjected in the marrow recipient. Blood. 1992; 80(10):2661-2667. (Clone-specific: Depletion, Flow cytometry, Functional assay, In vivo exacerbation). View Reference
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Wu L, Antica M, Johnson GR, Scollay R, Shortman K. Developmental potential of the earliest precursor cells from the adult mouse thymus. J Exp Med. 1991; 174(6):1617-1627. (Clone-specific: Cell separation, Depletion). View Reference
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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.