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PerCP-Cy5.5 Mouse Anti-Human CD80 (B7-1)
PerCP-Cy5.5 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 preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (Cat No. 552834; dashed line histogram) or PerCP-Cy5.5 Mouse Anti-Human CD80 (B7-1) antibody (Cat. No. 567437/567436; 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 FACSCelesta™ 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 preincubated with Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either PerCP-Cy5.5 Mouse IgG1, κ Isotype Control (Cat No. 552834; dashed line histogram) or PerCP-Cy5.5 Mouse Anti-Human CD80 (B7-1) antibody (Cat. No. 567437/567436; 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 FACSCelesta™ Cell Analyzer System and FlowJo™ software.

Product Details
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BD Pharmingen™
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 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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

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. 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. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  8. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  11. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567437 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.

567437 Rev. 1
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
567437 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
View All (6) View Less
567437 Rev. 1

Please refer to Support Documents for Quality Certificates


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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.