Bio plex pro mouse cytokine

Post-traumatic inflammation was measured using a multiplex enzyme-linked immunosorbent assay (ELISA) in diabetic animals treated with hBM-MSC regularly fed, starved, and EVs derived from hBM-MSC; diabetic fractured and non-diabetic fractured animals were used as sham (n = 3 to 6). Plasma concentrations of 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-17, eotaxin, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-γ(IFN-γ), keratinocyte-derived chemokine (KC, CXCL1), monocyte chemotactic protein 1(MCP-1), macrophage inflammatory protein-1α, 1β (MIP-1α and MIP-1β), regulated on activation, normal T cell expressed and secreted (RANTES) and tumor necrosis factor-α (TNF-α) were quantified using 50 μL of blood and the Bio-Plex Pro Mouse Cytokine (Bio-Rad Laboratories, Hercules, CA, USA). Data were analysed with the Luminex 200 Total System (Bio-Rad).

In experiment 2, the water intake, food intake, and body weight of mice were recorded daily. The right lung lobes of mice were collected to prepare 10% lung homogenates by using an electric homogenizer on ice, and the homogenates were stored at –80 °C for cytokine assays [33 (link)]. The levels of tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), IL-4, IL-5, IL-10, IL-12p70, interferon-γ (IFN-γ), and granulocyte–macrophage colony-stimulating factor (GM-CSF) in lung homogenates were detected by Luminex Liquid Chip Technology in accordance with the manufacturer’s instructions (Bio-Plex Pro™ Mouse Cytokine, Bio-Rad).

At Days 3 and 6, blood was collected and processed to isolate plasma; bronchoalveolar lavage (BAL) samples were collected and processed. Cytokine content in both types of samples was analyzed using the Bio-Plex Pro Mouse Cytokine, Chemokine, and Growth Factor Assays and the Bio-Plex 200 system (Bio-Rad Laboratories, Watford, UK) by following the manufacturer’s instructions with no modifications. Reference standards were included in duplicate for every analyte analyzed, in order to quantify cytokine and chemokine. Results from samples were interpolated into standard curves for each analyte. Standard diluent was used as a blank. The analytes quantified were as follows: IL-1β, IL-2, IL-6, IL-10, IL-12p70, IL-17, IFN-γ, GM-CSF, MCP-1, and TNF-α.