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Three-color image analysis of CD11c expression by cells within C57BL/6 mouse spleen. Mouse spleen cryosections (5 µm) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1× PBS, and stained with BD Horizon™ BV480 Hamster Anti-Mouse CD11c antibody (Cat. No. 565627, pseudo-colored green), Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669, pseudo-colored blue), and Alexa Fluor® 647 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 557869, pseudo-colored red). Slides were mounted with ProLong® Gold and the images were captured on a standard epifluorescence microscope. Acquired at 20× magnification.
BD Horizon™ BV480 Hamster Anti-Mouse CD11c
Regulatory Status Legend
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
Recommended Assay Procedures
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. 563794/566349).
For Immunofluorescence Applications:
The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480. For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter.
For epifluorescence microscopes with broad spectrum excitation sources, the recommended excitation and emission filters are 445/20 nm and 485/20 nm bandpass filters, respectively. For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging.
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.
- 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.
- BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
- ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The HL3 monoclonal antibody specifically binds to the integrin αx chain of gp150, 95 (CD11c/CD18). CD11c is expressed on dendritic cells, CD4- CD8+ intestinal intraepithelial lymphocytes (IEL) and some NK cells. It is upregulated on IEL and lymph-node T cells following in vivo activation. Cells of the monocyte/macrophage lineage have been reported to express low levels of CD11c. CD11c plays a role in binding of iC3b.
The antibody was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set. BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.
Development References (9)
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Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
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Burt BM, Plitas G, Stableford JA, et al. CD11c identifies a subset of murine liver natural killer cells that responds to adenoviral hepatitis. J Leukoc Biol. 2008; 84(4):1039-1046. (Clone-specific: Flow cytometry). View Reference
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Fagarasan S, Muramatsu M, Suzuki K, Nagaoka H, Hiai H, Honjo T. Critical roles of activation-induced cytidine deaminase in the homeostasis of gut flora. Science. 2002; 298(5597):1424-1427. (Clone-specific: Immunofluorescence). View Reference
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Huleatt JW, Lefrançois L. Antigen-driven induction of CD11c on intestinal intraepithelial lymphocytes and CD8+ T cells in vivo.. J Immunol. 1995; 154(11):5684-93. (Immunogen: Flow cytometry, Immunoprecipitation). View Reference
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Larson RS, Springer TA. Structure and function of leukocyte integrins. Immunol Rev. 1990; 114:181-217. (Biology). View Reference
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Maraskovsky E, Brasel K, Teepe M, et al. Dramatic increase in the numbers of functionally mature dendritic cells in Flt3 ligand-treated mice: multiple dendritic cell subpopulations identified. J Exp Med. 1996; 184(5):1953-1962. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Metlay JP, Witmer-Pack MD, Agger R, Crowley MT, Lawless D, Steinman RM. The distinct leukocyte integrins of mouse spleen dendritic cells as identified with new hamster monoclonal antibodies. J Exp Med. 1990; 171(5):1753-1771. (Biology). View Reference
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Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. (Clone-specific: Immunohistochemistry). View Reference
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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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.