Biorad bio plex software

The levels of various cytokines in the serum of mice were measured using a MILLIPLEX Map Human Cytokine/Chemokine kit (Millipore, Billerica, MA, USA) on a Luminex 200 system (Luminex, Austin, TX, USA), which employed xMAP technology, according to the manufacturer’s instructions. Briefly, wash buffer was added into each well of a plate, after which the buffer was removed, and premixed standard cocktails and serum samples were added. Beads coupled with antibodies against tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), vascular endothelial growth factor (VEGF), interleukin (IL)-2, IL-6, interferon-γ (IFN-γ), granulocyte colony stimulating factor (G-CSF), and matrix metalloproteinase-9 (MMP-9) were diluted in blocking buffer. Each sample was then incubated with the bead solution for 2 h at room temperature. Following one wash, a premixed detection antibody cocktail was added, and the plate was incubated for 1 h. The streptavidin–phycoerythrin mixture was added to each well, and then the beads were resuspended in PBS. The Luminex 200 platform coupled with BioRad Bio-Plex software (BioRad, Hercules, CA, USA) was used to measure the cytokine levels.