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BV786 Hamster Anti-Mouse CD69
BV786 Hamster Anti-Mouse CD69
Flow cytometric analysis of CD69 expression on stimulated mouse splenocytes. 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™ BV786 Hamster IgG1, λ1 Isotype Control (Cat. No. 563751; dashed line histogram) or BD Horizon BV786 Hamster Anti-Mouse CD69 antibody (Cat. No. 564683; solid line histogram).  The fluorescence histogram showing CD69 expression (or Ig Isotype control staining) was 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.
Flow cytometric analysis of CD69 expression on stimulated mouse splenocytes. 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™ BV786 Hamster IgG1, λ1 Isotype Control (Cat. No. 563751; dashed line histogram) or BD Horizon BV786 Hamster Anti-Mouse CD69 antibody (Cat. No. 564683; solid line histogram).  The fluorescence histogram showing CD69 expression (or Ig Isotype control staining) was 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
12515
AB_2738890
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 with BD Horizon™ BV786 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV786 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Cy is a trademark of GE Healthcare.
  7. BD Horizon Brilliant Violet 786 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. 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.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564683 Rev. 2
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.

564683 Rev. 2
Format Details
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BV786
The BD Horizon Brilliant Violet™ 786 (BV786) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an Ex Max of 407-nm and an acceptor dye with an Em Max at 786-nm.  BV786, driven by BD innovation, is designed to be excited by the violet laser and detected using a filter, centered near 785 nm (e.g. 780/60 nm bandpass filter).  Please ensure that your instrument’s configurations (lasers and filters) are appropriate for this dye.
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BV786
Violet 405 nm
407 nm
786 nm
564683 Rev.2
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: Flow cytometry). View Reference
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564683 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.