Human mcp 1 enzyme linked immunosorbent assay elisa kits

The MCP-1 levels in the plasma samples were analyzed using human MCP-1 enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems, Abingdon, UK) as previously described [56 (link)]. Briefly, the wells of an ELISA plate containing 0.1 mL of diluted samples were incubated at 37 °C for 2 h and, subsequently, 0.1 mL of anti-human MCP-1 antibody was added to each well. Each well was washed three times with 0.4 mL washing buffer, and then 0.2 mL of conjugate solution was added. Again, the wells were washed and loaded with 0.2 mL of TMB agent at 37 °C in a dark room for 30 min. For final processing, each well was filled with 0.1 mL of stop solution, and the amount of MCP-1 absorbance was quantitatively measured at 450 nm with a spectrophotometer and calibrated using human MCP-1 standards.

The MCP-1 levels in the plasma samples were analyzed using human MCP-1 enzyme-linked immunosorbent assay (ELISA) kits (R and D Systems, Abingdon, UK). The results were calculated from a standard curve generated by diluting a known amount of recombinant MCP-1 protein. Each standard or sample was assayed in duplicate.