Hsp2mag 63 k

MMPs and cytokines analysis was performed by using xMAP technology (Luminex 200, Luminex, Austin, TX, USA) to measure MMP-1, MMP-2, MMP-8, MMP-9, IL-1β, and IL-8. The Milliplex MAP multiplex assay Human MMP Panel 2 Magnetic Bead HMMP2MAG-55 K and HSP2MAG-63 K (Millipore, Billerica, MA, USA) kits were conducted in 96-well microplate format according to the manufacturer’s recommendations. Concentrations of MMPs were determined on the Bio-Plex Protein Array System (Bio-Rad, Hercules, CA, USA). Briefly, each of the bead solutions was transferred into a mixing vial and adjusted to a volume of 3 ml with bead diluents. Internal controls and standards were included in every assay. Following the addition of sample supernatants and beads, the resulting mixture was incubated overnight at 4 °C. The plate was read by Bio-Plex array reader with the Luminex 200 software system [20 (link)].

Immunoassay analysis was performed by using xMAP technology (Luminex 200; Luminex, Austin, TX, USA) to measure MMP‐1, MMP‐2, MMP‐8, MMP‐9, IL‐1β, and IL‐8. The Milliplex MAP multiplex assay Human MMP Panel 2 Magnetic Bead HMMP2MAG‐55K and HSP2MAG‐63K (Millipore, Billerica, MA, USA) kits were used. Analysis was conducted in 96‐well microplate format according to the manufacturer's recommendations. Concentrations of MMPs were determined on the Bio‐Plex Protein Array System (Bio‐Rad, Hercules, CA, USA). Briefly, each of the bead solutions was transferred into a mixing vial and adjusted to a volume of 3 ml with bead diluents. Internal controls and standards were included in every assay. Following the addition of sample supernatants and beads, the resulting mixture was incubated overnight at 4°C. The plate was scanned using a Bio‐Plex array reader with the Luminex 200 software system.