Q view software program

Levels of human cytokines IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17, IL-23 IFNγ and TNFα and chemokines GROα, Eotaxin, I-309, IP-10, MCP-1 and MCP-2 were detected using Q-Plex technology (Quansys Biosciences, Logan, UT, USA) according to the manufacturer’s instructions. Readouts were obtained with a Quansys Imager (Quansys Biosciences) and results analyzed using the Q-View Software program (Quansys Biosciences). A human sL-Selectin Instant ELISA (eBioscience) was also performed according to the manufacturer’s recommended protocol, with the exception that culture supernatants were diluted 1:10, of which 50 μL was added to provide plates in duplicate.

Levels of human cytokines were detected using Q-Plex technology (Quansys Biosciences, Logan, UT, USA) according to manufacturer's instructions. Briefly, 30 μL of each sample (duplicate) was diluted 1:2 in diluent buffer, and a total volume of 50 μL of diluted sample and control standards was added to a 96-well v-bottom binding plate. Plates were then incubated for 1 h on a shaker at room temperature in the dark. Following incubation, plates were washed and a volume of 50 μL of detection mix was added to each well, and incubated for 1 h. Next, plates were washed and a volume of 50 μL streptavidin HRP was added to each well and incubated for additional 15 min. Finally, plates were washed and a volume of 50 μL substrate mix was added to each well. Readouts were obtained with a Quansys Imager (Quansys Biosciences, Logan, UT, USA) and results analyzed using the Q-View Software program (Quansys Biosciences).