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R718 Mouse Anti-Human CD80 (B7-1)
R718 Mouse Anti-Human CD80 (B7-1)

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.

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.

Product Details
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BD Horizon™
B7; B7-1; B7.1; BB1; CD28LG
Human (QC Testing)
Mouse IgG1, κ
Not Reported
Flow cytometry (Routinely Tested)
5 µl
VI CD80.1
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 to the dye under optimum conditions and unreacted dye was removed.

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

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567441 Rev. 1
Antibody Details
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2D10.4

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.

567441 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567441 Rev.1
Citations & References
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Development References (6)

  1. 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
  2. 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.
  3. 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
  4. 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
  5. 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
  6. 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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567441 Rev. 1

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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.