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Flow cytometric analysis of CD80 (B7-1) expression on human Raji cells. Cells from the human Raji (Burkitt's B cell lymphoma, ATCC CCL-86) cell line were stained with either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat No. 566928; dashed line histogram) or BD Horizon™ R718 Mouse Anti-Human CD80 (B7-1) antibody (Cat. No. 567442/567441 solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing CD80 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) Raji cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Horizon™ R718 Mouse Anti-Human CD80 (B7-1)
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
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- 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.
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The 2D10.4 monoclonal antibody, also known as 2D10, specifically recognizes CD80 which is otherwise known as Activation B7-1 antigen (B7-1), B7.1, or B7. This ~60 kDa single-pass type I transmembrane glycoprotein is encoded by CD80 (CD80 molecule) that belongs to the immunoglobulin gene superfamily (IgSF). CD80 (B7-1) is comprised of an extracellular region with an IgV-like domain followed by an IgC-like domain, a transmembrane region and a cytoplasmic tail. It is variably expressed on activated B cells, activated CD4+ and CD8+ T cells, macrophages, and dendritic cells. CD80 (B7-1) serves as a ligand for the CD28 and CD152 (CTLA-4) coreceptors that are differentially expressed on T cells. CD80 (B7-1) engagement of CD28 costimulates T cells activated through their TCR-CD3 complex whereas its interaction with CD152 (CTLA-4) generates an inhibitory signal for T cell responses.
The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.
Development References (6)
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Battifora M, Pesce G, Paolieri F, et al. B7.1 costimulatory molecule is expressed on thyroid follicular cells in Hashimoto's thyroiditis, but not in Graves' disease.. J Clin Endocrinol Metab. 1998; 83(11):4130-9. (Clone-specific: Flow cytometry, Functional assay, Immunocytochemistry, Immunohistochemistry). View Reference
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Jones M, Mason DY. CD80 Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:186-187.
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Mielle J, Audo R, Hahne M, et al. IL-10 Producing B Cells Ability to Induce Regulatory T Cells Is Maintained in Rheumatoid Arthritis. Front Immunol. 2018; 9:961. (Clone-specific: Flow cytometry). View Reference
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Orbach A, Rachmilewitz J, Shani N, et al. CD40·FasL and CTLA-4·FasL fusion proteins induce apoptosis in malignant cell lines by dual signaling.. Am J Pathol. 2010; 177(6):3159-68. (Clone-specific: Flow cytometry). View Reference
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Werz O, Gerstmeier J, Libreros S, et al. Human macrophages differentially produce specific resolvin or leukotriene signals that depend on bacterial pathogenicity. Nature Commun. 2018; 9(1):59. (Clone-specific: Flow cytometry). View Reference
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van der Merwe PA, Bodian DL, Daenke S, Linsley P, Davis SJ. CD80 (B7-1) binds both CD28 and CTLA-4 with a low affinity and very fast kinetics.. J Exp Med. 1997; 185(3):393-403. (Clone-specific: Western blot). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.