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Pe conjugated cd86

Manufactured by BioLegend
Sourced in United States

PE-conjugated CD86 is a laboratory reagent that consists of the CD86 antibody conjugated to the fluorescent dye Phycoerythrin (PE). CD86 is a cell surface protein that is expressed on antigen-presenting cells and plays a role in T cell activation and immune response. The PE-conjugated CD86 antibody can be used to identify and study CD86-expressing cells in research applications.

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3 protocols using pe conjugated cd86

1

Flow Cytometric Analysis of ARPE-19 Cell Surface Markers

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The cultured ARPE-19 cells were analyzed using flow cytometry. The cells were harvested by treatment with 0.25% trypsin- 0.5% EDTA (Gibco, Carlsbad, CA, USA), washed with 1X PBS, and the cell pellet was resuspended in 2% fetal bovine serum in 1X PBS for 1 h at room temperature.
The identification of surface molecules was carried out by incubating the cells with human monoclonal APC-conjugated anti-toll like receptor 4 (TLR4), FITC-conjugated CD14, and PE-conjugated CD86 antibodies (all from Biolegend, San Diego, CA, USA) for 20 min at 4 °C, according to the manufacturer’s recommendations. Cells were washed with 1X PBS and fixed with 4% paraformaldehyde (Sigma-Aldrich, Inc.) for 20 min at room temperature, washed, and acquired with a BD FACSVerse Flow Cytometer (BD Biosciences, San Jose, CA, USA). The results were analyzed using Flow Jo ver. 10 (Beckton Dickinson, East Rutherford, NJ, USA).
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2

Flow Cytometric Analysis of Dendritic Cell Maturation

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The isolated dendritic cells, either from uninfected control mice, E. hellem infected mice or LPS treated mice were all subjected to flow cytometer and analyzed for cell maturation. In brief, the DCs were firstly washed with 1× PBS/0.3% BSA, and then stained with FITC-conjugated anti-CD40 and PE-conjugated CD86 (BioLegend, San Diego, CA, USA) for 30 min at 4 °C. CD40+/CD86+ DCs were considered as more matured DCs. Flow cytometry analysis was performed by a FACSCanto II flow cytometer (BD Biosciences) and data were collected and analyzed with FACSDiva software (v6.1.2).
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3

Immunophenotypic Characterization of Cells

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Cells were dissociated from 24-well culture dishes using Accutase® cell detachment solution (Sigma), washed in dPBS containing 5 % FBS, 5 mM Ethylenediaminetetraacetic acid (EDTA) and 0.1 % sodium azide (Sigma) and probed for HLA-ABC, HLA-DR/DP/DQ, CD80, CD86, PDL1 and/or CXCR3 using the following antibodies: Pacific Blue-conjugated HLA-ABC (Biolegend) Brilliant Blue 515-conjugated HLA-DR/DP/DQ, phycoerythrin (PE)-conjugated CD86, allophycocyanin-conjugated PDL1, PE/Cy7-conjugated CD80 or PE/Cy7-conjugated CXCR3 (Biolegend). All antibodies were purchased from BD Biosciences unless otherwise indicated and used at manufacturer’s recommended dilutions. Manufacturer’s recommended isotype controls were used as negative controls for all antibodies used. A total of 10,000 events per sample were acquired using a BD LSR II (BD Biosciences), and data were analyzed using FlowJo data analysis software (FLOWJO LLC, Ashland, OR).
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