Quantikine elisa human kits

The relative concentration of plasma cytokine expression was measured by ELISA using the Human ICAM-1/CD54, IL-8/CXCL8, MIF, IL-6 and C-Reactive Protein/CRP via the Quantikine ELISA Human kits (R&D Systems, Minneapolis, MN, USA). Kits were used according to manufacturer's instructions. Briefly, plates were coated with monoclonal anti-human cytokine capture antibody and blocked with 1% bovine serum albumin. Plasma samples, in triplicate, were incubated for two hours at room temperature together with recombinant human cytokine standards. Bound cytokine was detected with anti-human cytokine horseradish peroxidase secondary antibody and tetramethylbenzidine substrate. The concentration of cytokine per sample was calculated using a four-parameter logistic curve fit of the standard concentrations. All data were expressed as mean ± S.D. and for comparisons of the means, the student t-test was used. p values less than 0.05 were considered statistically significant.