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2 protocols using gro α

1

Multiplex Cytokine Quantification in BALF

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Chemokine and cytokine concentrations in BALF supernatants were quantified using Bio-Plex multiplex bead-based assays by means of Bio-Plex Pro™ Human Cytokine 27-Plex Immunoassay and three individual assays for human VCAM-1, growth regulated protein-α (GRO-α) and hepatocyte growth factor (HGF) (Bio-Rad Laboratories, Marnes-la-Coquette, France). For each cytokine calibration curve, eight standards were used. Data acquisition and analysis were completed using the Bio-Plex® 200 System with workstation Bio-Plex Manager™ software Version 5.0 (Bio-Rad Laboratories). Each condition was performed in duplicate.
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2

Cytokine Profiling of LPS-Stimulated Monocytes

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LPS isolated from Escherichia coli (0111:B4) was purchased from Sigma–Aldrich (St. Louis, MO, USA). Multiplex beads for: IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12 (p70), IL-17, G-CSF, GM-CSF, IFN-γ, MCP-1, MIP-1α/1β, RANTES, TNF-α, VEGF, IP-10, and GRO-α were purchased from Bio-Rad Laboratories (Hercules, CA, USA). TGFβ was purchased from Peprotech (Rocky Hill, NJ, USA) and neutralizing TGFβ Ab was purchased from R&D (Minneapolis, MN, USA). For flow cytometry PE-conjugated anti human-CD14 and FITC-conjugated anti human-CD16 Abs purchased from eBioscience (San Diego, CA, USA) were used at a 1:50 dilution.
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