Human sflt 1

Enzyme-linked immunosorbent assays (ELISAs) were performed using commercial kits. Human EGFL7 kits were purchased from Cusabio Biotech (College Park, Maryland, USA). Human sFLT-1, free PlGF, and sENDOGLIN kits were purchased from R&D Systems (Minneapolis, Minnesota, USA). Assays were performed according to the manufacturer’s instructions in the Department of Biomedicine and Prevention, University of Rome Tor Vergata, Italy. Data on spike/recovery, inter- and intra- assay precision were provided by the manufacturer. In details, intra-assay precision was indicated as coefficient of variability (CV) % and calculated as < 8% by testing three samples of known concentration twenty times on one plate. Inter-assay precision was reported as CV% < 10% and it was obtained by testing three samples of known concentration in twenty assays. To confirm these data, we tested five different plasma samples in at least three different assays and obtained comparable results. In each ELISA, all samples and standards have been tested in duplicate; duplicates always gave similar values of optical density (OD).