Multiplexed Cytokine Profiling of Cell Cultures

After 24-h incubation with samples in 96-well plates, the following cytokines production from each well containing 10,000 cells was analyzed: interleukin (IL)-6; IL-10, tumor necrosis factor (TNF)-α; granulocyte colony-stimulating factor (G-CSF); granulocyte macrophage colony-stimulating factor (GM-CSF); lipopolysaccharide-induced CXC chemokine (LIX; CXCL5); monocyte chemotactic activating factor (MCP)-1; macrophage inflammatory protein (MIP)-1α, MIP-1β, MIP-2, RANTES (CCL5; regulated on activation, normal T cell expressed and secreted), vascular endothelial growth factor (VEGF); interferon gamma-induced protein 10 (IP-10; CXCL10); and leukemia inhibitory factor (LIF). Cytokines were measured using a Luminex assay based on xMAP technology with multiplex cytokine assay kits and a Bio-Plex 200 suspension array system (Bio-Rad, Hercules, CA, USA), as described previously [4 (link),5 (link),6 (link),7 (link),8 (link),9 (link)]. The assay used in this experiment was designed for the multiplexed quantitative measurement of multiple cytokines in a single well, using as little as 50 µL of cell culture supernatant. Baicalein, a well-known anti-inflammatory flavonoid, was used as a positive control.

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Kim Y.J., Lee J.Y., Kim H.J., Kim D.H., Lee T.H., Kang M.S, & Park W. (2018). Anti-Inflammatory Effects of Angelica sinensis (Oliv.) Diels Water Extract on RAW 264.7 Induced with Lipopolysaccharide. Nutrients, 10(5), 647.

Publication 2018

multiplex cytokine assay kits Bio-Plex 200 suspension array system

Anti inflammatory Assay Baicalein Ccl5 Cell culture Chemotactic factor Cxc chemokine Cxcl5 Cytokines Flavonoid Gm csf Granulocyte colony stimulating factor Interferon gamma Interleukin 10 Interleukin 6 Leukemia inhibitory factor Lipopolysaccharide Macrophage inflammatory protein 1α Macrophage inflammatory protein 1β Macrophage inflammatory protein 2 Mcp 1 Monocyte Normal cell Protein Rantes Tumor necrosis factor α Vascular endothelial growth factor

Corresponding Organization : Gachon University

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Protocol cited in 3 other protocols

Variable analysis

independent variables

Baicalein, a well-known anti-inflammatory flavonoid

dependent variables

Interleukin (IL)-6
IL-10
Tumor necrosis factor (TNF)-α
Granulocyte colony-stimulating factor (G-CSF)
Granulocyte macrophage colony-stimulating factor (GM-CSF)
Lipopolysaccharide-induced CXC chemokine (LIX; CXCL5)
Monocyte chemotactic activating factor (MCP)-1
Macrophage inflammatory protein (MIP)-1α, MIP-1β, MIP-2
RANTES (CCL5; regulated on activation, normal T cell expressed and secreted)
Vascular endothelial growth factor (VEGF)
Interferon gamma-induced protein 10 (IP-10; CXCL10)
Leukemia inhibitory factor (LIF)

control variables

10,000 cells per well
24-h incubation

positive control

Baicalein, a well-known anti-inflammatory flavonoid

negative control

Not explicitly mentioned

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