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 BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either PE Mouse IgG1, κ Isotype Control (Cat No. 562438, dashed line histogram) or PE Mouse Anti-Human CD80 (Cat. No. 554680; solid line histogram). 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 human Raji cells. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software.
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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 preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220) and stained with either PE Mouse IgG1, κ Isotype Control (Cat No. 562438, dashed line histogram) or PE Mouse Anti-Human CD80 (Cat. No. 554680; solid line histogram). 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 human Raji cells. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Cell Analyzer System and FlowJo™ software.
Product Details
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
AB_2869994
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Human BD Fc Block™ RUO
Size
50 µg
Cat No.
564219
Human BD Fc Block™ RUO
Size
0.25 mg
Cat No.
564220
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
566992 Rev. 1
Antibody Details
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.
566992 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566992 Rev.1
Citations & References
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
566992 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.
