Bio plex mouse cytokine 23 plex panel kit

Serum was collected from blood drawn by cardiac puncture at the end of the treatment. Explants from the proximal and distal colons of treatment and control groups (n = 3) were cultured overnight in RPMI 1640. Culture supernatants were measured for Nitrite by the Griess reaction method (Sigma G4410) as an index of Nitric Oxide generation.
Cytokine concentrations in neat culture supernatants and serum were determined using mouse Bio-Plex mouse cytokine 23-plex panel kit (Bio-Rad #M60009RDPD) and analysed using Luminex 200 (Bio-Rad) and Bio-Plex Manager software (Bio-Rad). IL18 was determined by 5 times diluted supernatant measured by a mouse IL18 ELISA kit (7625, R&D Systems). The most significantly altered cytokines are presented as pg per g of tissue.

Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured using a dry chemistry analyzer (FUJI DRI-CHEM 7000V, FUJIFILM, Tokyo, Japan). To indirectly quantify endotoxin levels, lipopolysaccharide (LPS)-binding protein (LBP) concentrations were measured by ELISA according to the manufacturer’s guidelines (Guduo, Shanghai, China) because of the well-documented inaccuracies inherent to direct endotoxin measurements (Tremellen et al., 2017 (link)). Serum cytokine levels were measured using a Bio-Plex Mouse Cytokine 23-Plex Panel kit (Bio-Rad, Hercules, CA, United States) according to the manufacturer’s instructions. The following mouse cytokine levels were analyzed for each sample: interleukin (IL)-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12 p40, IL-12 p70, IL-13, IL-17α, eotaxin, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), gamma interferon (IFN-γ), keratinocyte-derived chemokine (KDC), macrophage chemoattractant protein 1 (MCP)-1, macrophage inflammatory protein-1α (MIP)-1α, MIP-1β, regulated upon activation normal T-cell expressed and secreted (RANTES), and tumor necrosis factor (TNF)-α.