Anti cd86 brilliant violet 421

Manufactured by BioLegend

Sourced in United States

Anti-CD86-Brilliant Violet 421 is a fluorescently labeled antibody product from BioLegend. It is designed to detect the expression of CD86, a costimulatory molecule, on the surface of cells. The Brilliant Violet 421 fluorophore is used to label the antibody, enabling flow cytometric analysis of CD86-positive cells.

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3 protocols using anti cd86 brilliant violet 421

Characterizing B-cell CD39 and CD73 Expression

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Peripheral blood mononuclear cells (PBMCs) were isolated from freshly heparinized blood. To characterize the expression profiles of CD39 and CD73 on B-cells, PBMCs or intra-hepatic lymphocytes were stained with a cocktail of monoclonal antibodies (mAbs): anti-CD45-Allophycocyanin-H7 (clone 2D1, BD Biosciences, Franklin Lakes, NJ, USA), anti-CD19-Phycoerythrin-Cy7 (clone SJ25C1, BD Biosciences), anti-CD39-Fluorescein isothiocyanate (clone TU66, Biolegend, San Diego, CA, USA), anti-CD73-Brilliant violet 421 (clone AD2, Biolegend), anti-CD10-Phycoerythrin (clone HI10a, BD Biosciences), anti-CD27-Brilliant violet 510 (clone L128, Biolegend), and anti-CD21-Allophycocyanin (clone B-ly4, eBioscience, San Diego, CA, USA).
To detect the B-cell-activation phenotypes, cells were stained with the following surface mAbs: anti-CD19-PerCP-Cy5.5 (clone HIB19, BD Biosciences), anti-CD69-Fluorescein isothiocyanate (clone FN50, BD Biosciences), anti-CD71-Phycoerythrin-Cy7 (clone CY1G4, Biolegend), anti-CD80-Allophycocyanin (clone 2D10, Biolegend), and anti-CD86-Brilliant violet 421 (clone IT2.2, Biolegend). Flow-cytometer acquisition was performed on an LSR II instrument (BD Biosciences) and data were analysed using FlowJo (Version 7.6.1; TreeStar) software.

Zhou S.N., Zhang N., Liu H.H., Xia P., Zhang C., Song J.W., Fan X., Shi M., Jin L., Zhang J.Y, & Wang F.S. (2020). Skewed CD39/CD73/adenosine pathway contributes to B-cell hyperactivation and disease progression in patients with chronic hepatitis B. Gastroenterology Report, 9(1), 49-58.

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Profiling Immune Cell Activation

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The harvested tumors were cut into small pieces, digested at 37 °C for 1 h, and filtered through 70 μm cell strainers to obtain single cell suspensions. After blocking with FcγIII/IIR-blocking Ab, the cells were stained with fluorescence-labeled antibodies at a dilution of 1:200. For macrophage re-polarization, the cells were stained with anti-CD86-Brilliant Violet 421™ (BioLegend, Clone: GL-1, Catalog: B318824) and anti-CD206-PE-CYN7 (Thermo, Clone: MR6F3, Catalog: 25-2061-82). For DCs maturation, the cells were stained with Anti-CD11c-APC/Cy7® (Abcam, Clone: 3.9, Catalog: ab272330), and anti-CD86-Brilliant Violet 421™. For T cells activation, the cells were stained with anti-CD8a-PE (BD Pharmingen, Clone: 53-6.7, Catalog: 553032), anti-CD4-Percp/Cy™5.5 (BD Pharmingen, Clone: RM4-5, Catalog: 550954) and anti–IFN–γ-FITC (BD, Clone: XMG1.2, Catalog: 554411). Afterward, the samples were analyzed by flow cytometry (FACSVerse, BD, USA).

Meng Y., Huang J., Ding J., Zhou H., Li Y, & Zhou W. (2024). Mn-phenolic networks as synergistic carrier for STING agonists in tumor immunotherapy. Materials Today Bio, 26, 101018.

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Flow Cytometry Immunophenotyping Protocol

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Following stimulation for flow cytometry experiments, cells were fixed with 4% para-formaldehyde for 10 min before being washed 3 times with PBS and blocked/quenched with 5% bovine serum + 0.1 mM glycine overnight at 4°C. Cells were stained with 1 μg/ml of anti-BTLA AlexaFluor 647 (RRID AB_2650979; BioLegend Cat. No. 344519), anti-CTLA4 PE (RRID AB_10645522; BioLegend Cat. No. 349905), anti-CD86 Brilliant Violet 421 (RRID AB_10899582; BioLegend Cat. No. 305425), anti-CD69 APC (RRID AB_314844; BioLegend Cat. No. 310909), or anti-CD25 FITC (RRID AB_314273; BioLegend Cat. No. 302603) for 1 h at room temperature then washed 3 times with PBS before being analysed using a FACSCanto II™ flow cytometer (BD Biosciences). Data were analysed using FlowJo version 8.8.7. Staining was performed in three independent experiments with cells from different donors.

Felce S.L., Farnie G., Dustin M.L, & Felce J.H. (2021). RNA-Seq analysis of early transcriptional responses to activation in the leukaemic Jurkat E6.1 T cell line. Wellcome Open Research, 5, 42.

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