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      BV605 Hamster Anti-Mouse CD69
      BV605 Hamster Anti-Mouse CD69
      Flow cytometric analysis of CD69 expression on stimulated mouse splenocytes. C57BL/6 mouse splenic leucocytes were stimulated for 5 hours at 37°C with Phorbol 12-Myristate 13-Acetate (PMA, 10 ng/mL; Sigma-Aldrich, Cat. No. P-8139). The stimulated cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BV605 Hamster IgG1, λ1 Isotype Control (Cat. No. 563054; dashed line histogram) or BD Horizon™ BV605 Hamster Anti-Mouse CD69 antibody (Cat. No. 563290; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable stimulated leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      BV605 Hamster Anti-Mouse CD69
      Flow cytometric analysis of CD69 expression on stimulated mouse splenocytes. C57BL/6 mouse splenic leucocytes were stimulated for 5 hours at 37°C with Phorbol 12-Myristate 13-Acetate (PMA, 10 ng/mL; Sigma-Aldrich, Cat. No. P-8139). The stimulated cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with either BD Horizon™ BV605 Hamster IgG1, λ1 Isotype Control (Cat. No. 563054; dashed line histogram) or BD Horizon™ BV605 Hamster Anti-Mouse CD69 antibody (Cat. No. 563290; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable stimulated leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Horizon™
      VEA; Very Early Activation Antigen; AIM; Activation Induced Molecule
      Mouse (QC Testing)
      Armenian Hamster IgG1, λ3
      Mouse Dendritic Epidermal T Cell Line Y245
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      AB_2738120
      Aqueous buffered solution containing ≤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 with BD Horizon™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.
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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      4. 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.
      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. 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.
      8. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
      9. CF™ is a trademark of Biotium, Inc.
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      563290 Rev. 3
      Antibody Details
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      H1.2F3

      The H1.2F3 monoclonal antibody specifically binds to CD69 (Very Early Activation antigen), an 85 kDa disulfide-linked homodimer of differentially glycosylated subunits. CD69 is a C-type lectin, most closely related to the NKR-P1 and Ly-49 NK cell-activation molecules. Its expression is rapidly induced upon activation of lymphocytes (T, B, NK, and NK-T cells), neutrophils, and macrophages. CD69 is expressed also on thymocytes that are undergoing positive selection; its role in that process is unclear. H1.2F3 mAb augments PMA-induced T-cell stimulation and IFN-γ-induced macrophage stimulation. IL-2-activated NK cells express CD69, and H1.2F3 mAb induces redirected lysis of FcR-bearing target cells by NK 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).

      563290 Rev. 3
      Format Details
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      BV605
      The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV605
      Violet 405 nm
      407 nm
      605 nm
      563290 Rev.3
      Citations & References
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      Development References (15)

      1. Bendelac A, Matzinger P, Seder RA, Paul WE, Schwartz RH. Activation events during thymic selection. J Exp Med. 1992; 175(3):731-742. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
      2. Gabor MJ, Godfrey DI, Scollay R. Recent thymic emigrants are distinct from most medullary thymocytes. Eur J Immunol. 1997; 27(8):2010-2050. (Clone-specific: Flow cytometry). View Reference
      3. Karlhofer FM, Yokoyama WM. Stimulation of murine natural killer (NK) cells by a monoclonal antibody specific for the NK1.1 antigen. IL-2-activated NK cells possess additional specific stimulation pathways. J Immunol. 1991; 146(10):3662-3673. (Clone-specific: Induction). View Reference
      4. Keefe R, Dave V, Allman D, Wiest D, Kappes DJ. Regulation of lineage commitment distinct from positive selection. Science. 1999; 286(5442):1149-1153. (Biology). View Reference
      5. Lauzurica P, Sancho D, Torres M, et al. Phenotypic and functional characteristics of hematopoietic cell lineages in CD69-deficient mice. Blood. 2000; 95(7):2312-2320. (Clone-specific: Flow cytometry). View Reference
      6. Marzio R, Jirillo E, Ransijn A, Mauel J, Corradin SB. Expression and function of the early activation antigen CD69 in murine macrophages. J Leukoc Biol. 1997; 62(3):349-355. (Clone-specific: Activation). View Reference
      7. Merkenschlager M, Graf D, Lovatt M, Bommhardt U, Zamoyska R, Fisher AG. How many thymocytes audition for selection. J Exp Med. 1997; 186(7):1149-1158. (Clone-specific: Flow cytometry). View Reference
      8. Nishimura T, Kitamura H, Iwakabe K, et al. The interface between innate and acquired immunity: glycolipid antigen presentation by CD1d-expressing dendritic cells to NKT cells induces the differentiation of antigen-specific cytotoxic T lymphocytes. Int Immunol. 2000; 12(7):987-994. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
      9. Punt JA, Suzuki H, Granger LG, Sharrow SO, Singer A. Lineage commitment in the thymus: only the most differentiated (TCRhibcl-2hi) subset of CD4+CD8+ thymocytes has selectively terminated CD4 or CD8 synthesis. J Exp Med. 1996; 184(6):2091-2099. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
      10. Sobel ES, Yokoyama WM, Shevach EM, Eisenberg RA, Cohen PL. Aberrant expression of the very early activation antigen on MRL/Mp-lpr/lpr lymphocytes. J Immunol. 1993; 150(2):673-682. (Clone-specific: Stimulation). View Reference
      11. Wilkinson RW, Anderson G, Owen JJ, Jenkinson EJ. Positive selection of thymocytes involves sustained interactions with the thymic microenvironment. J Immunol. 1995; 155(11):5234-5240. (Clone-specific: Cell separation, Flow cytometry). View Reference
      12. Yokoyama WM, Koning F, Kehn PJ, et al. Characterization of a cell surface-expressed disulfide-linked dimer involved in murine T cell activation. J Immunol. 1988; 141(2):369-376. (Immunogen: Flow cytometry, Immunoprecipitation, Stimulation). View Reference
      13. Yokoyama WM, Maxfield SR, Shevach EM. Very early (VEA) and very late (VLA) activation antigens have distinct functions in T lymphocyte activation. Immunol Rev. 1989; 109:153-176. (Clone-specific: Flow cytometry, Stimulation). View Reference
      14. Ziegler SF, Levin SD, Johnson L, et al. The mouse CD69 gene. Structure, expression, and mapping to the NK gene complex. J Immunol. 1994; 152(3):1228-1236. (Clone-specific: Flow cytometry). View Reference
      15. Ziegler SF, Ramsdell F, Alderson MR. The activation antigen CD69. Stem Cells. 1994; 12(5):456-465. (Biology). View Reference
      View All (15) View Less
      563290 Rev. 3

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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.

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