Human magnetic luminex screening assay elisa

Cell culture supernatants were assayed by Human Magnetic Luminex Screening Assay ELISA (R&D Systems, Minneapolis, MN, USA) to measure the concentration of inflammatory factors monocyte chemoattractant protein (MCP)-1, interleukin (IL)-6, IL-8, regulated upon activation, normal T cell expressed and presumably secreted (RANTES), and intercellular adhesion molecule (ICAM)-1. EA.hy926 cells were incubated with the FAs in 96-well flat-bottomed plates (Corning TM, Corning, NY) (1 × 10⁴ cells/100 µL per well) for 48 h and then incubated with TNF-α for a further 24 h. Before the supernatants of each well were collected and stored at −80 °C until analysis, the cells were checked under the microscope. Assays were conducted in accordance with the instructions from the manufacturer. Plates were analysed on a calibrated Bio-Plex 200 analyser using Bio-Plex software (version 6.1, Bio-Rad Laboratories Inc., Berkeley, CA, USA). Lower limits of detection (pg/mL) were IL-6, 1.7; IL-8, 1.8; MCP-1, 9.9; RANTES, 1.8; ICAM-1, 87.9. Due to differences in the ranges of fluorescence values among experiments, the results are presented as % of control.

Cell culture supernatants were assayed by Human Magnetic Luminex Screening Assay ELISA (R&D Systems, Minneapolis, MN, USA) to measure the concentration of inflammatory factors monocyte chemoattractant protein (MCP)-1, interleukin (IL)-6, IL-8, regulated upon activation, normal T cell expressed and presumably secreted (RANTES), and intercellular adhesion molecule (ICAM)-1. EA.hy926 cells were incubated with the FAs in 96 well plates (1 × 10⁴ cells/100 µL per well) for 48 h and then incubated with TNF-α for a further 24 h. Before the supernatants of each well were collected and stored at −80 °C until analysis, the cells were checked under the microscope. Assays were conducted in accordance with the instructions from the manufacturer. Plates were analysed on a calibrated Bio-Plex 200 analyser using Bio-Plex software (version 6.1, Bio-Rad Laboratories Inc., Berkeley, CA). Lower limits of detection (pg/mL) were IL-6, 1.7; IL-8, 1.8; MCP-1, 9.9; RANTES, 1.8; ICAM-1, 87.9. Due to differences in the ranges of fluorescence values among experiments, the results are presented as % of control.

Cell culture supernatants were assayed using the Human Magnetic Luminex Screening Assay ELISA (R&D Systems, Minneapolis, MN, USA) to measure the concentrations of monocyte chemoattractant protein (MCP)-1, interleukin (IL)-6, IL-8, regulated upon activation, normal T cell expression and presumably secreted (RANTES) and intercellular adhesion molecule (ICAM)-1. The EA.hy926 cells were incubated with the FAs in 96-well flat-bottomed plates (Corning Corning, NY, USA) (1 × 10⁴ cells/100 µL per well) for 48 h and then without FAs for 24 h. Supernatants were collected and stored at −80 °C until analysis. Assays were conducted in accordance with the manufacturer’s instructions. Plates were analysed on a calibrated Bio-Plex 200 analyser using Bio-Plex software (version 6.1, Bio-Rad Laboratories Inc., Berkeley, CA, USA). Lower limits of detection (pg/mL) were IL-6, 1.7; IL-8, 1.8; MCP-1, 9.9; RANTES, 1.8; ICAM-1, 87.9. Due to the differences in the ranges of fluorescence values among the experiments the results are presented as % of control.