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Flow cytometric analysis of CD80 on resting or stimulated mouse splenocytes. Freshly isolated (Left Panel) or 72-hour lipopolysaccharide (LPS)-stimulated mouse splenic leucocytes (Right Panel) 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™ BUV737 Hamster IgG2, κ Isotype Control (Cat. No. 564671; dashed line histograms) or BD Horizon BUV737 Hamster Anti-Mouse CD80 antibody (Cat. No. 564670; solid line histograms). The fluorescence histograms showing CD80 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable resting or activated leucocytes as indicated. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ BUV737 Hamster Anti-Mouse CD80
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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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
Companion Products
The 16-10A1 monoclonal antibody specifically recognizes CD80 (B7-1). This member of the Ig superfamily, like CD86 (B7-2), can bind to either CD28 or CD152 (CTLA-4) and provide either costimulatory or coinhibitory signals to T cells, respectively. CD80 is constitutively expressed on dendritic cells, monocytes, and peritoneal macrophages as well as by activated B cells and T cells. The 16-10A1 antibody blocks binding of CTLA-4 Ig to CD80 as well as T-cell activation by Con A-elicited peritoneal exudate cells or CD80-transfected cell lines. However, the 16-10A1 antibody alone is not able to block T-cell activation by antigen-presenting cells. The 16-10A1 antibody may reportedly block the binding of another CD80-specific antibody, clone 1G10. In addition, the 16-10A1 antibody may crossreact with an activation antigen expressed on IFN-γ-activated alveolar macrophages of the dog.
The antibody was conjugated to BD Horizon BUV737 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 737-nm. BD Horizon Brilliant BUV737 can be excited by the ultraviolet laser (355 nm) and detected with a 740/35 filter. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into channels detecting Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).
Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV737 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV737 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
Development References (5)
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Bluestone JA. New perspectives of CD28-B7-mediated T cell costimulation. Immunity. 1995; 2(6):555-559. (Biology). View Reference
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Boussiotis VA, Gribben JG, Freeman GJ, Nadler LM. Blockade of the CD28 co-stimulatory pathway: a means to induce tolerance. Curr Opin Immunol. 1994; 6(5):797-807. (Biology). View Reference
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Hathcock KS, Laszlo G, Pucillo C, Linsley P, Hodes RJ. Comparative analysis of B7-1 and B7-2 costimulatory ligands: expression and function. J Exp Med. 1994; 180(2):631-640. (Biology). View Reference
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Razi-Wolf Z, Freeman GJ, Galvin F, Benacerraf B, Nadler L, Reiser H. Expression and function of the murine B7 antigen, the major costimulatory molecule expressed by peritoneal exudate cells. Proc Natl Acad Sci U S A. 1992; 89(9):4210-4214. (Immunogen: Blocking, Immunoprecipitation). View Reference
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Sojka DK, Donepudi M, Bluestone JA, Mokyr MB. Melphalan and other anticancer modalities up-regulate B7-1 gene expression in tumor cells. J Immunol. 2000; 164(12):6230-6236. (Biology). 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.
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