Human pon1 elisa kit

PON1 concentration was analyzed with the Human PON1 ELISA kit from Elabscience^® (Houston, TX, USA). PON1 activity was analyzed by the rate of hydrolysis of phenylacetate at 280 nm, in a 9 mM Tris-HCl buffer, pH 8.0, and supplemented with 0.9 mM CaCl2, as previously described [4 (link)]. The specific activity of PON1 was calculated as the ratio between PON1 activity and concentration. CCL2, IL-10, IL-4 and INF- were quantified with ABTS ELISA Development kits (Peprotech, London, UK). Transforming growth factor beta 1 (TGF-β1) and TNF-α concentrations were measured by Proteintech ELISA kits (Manchester, United Kingdom). CD4+ and CD8+ cells were quantified by flow citometry (FACSCalibur^TM analyzer, Beckton & Dickinson, Franklin Lakes, NJ, USA). D-Dimer was analyzed in a Sysmex CS-2100i^TM analyzer (Sysmex GmbH, Norderstedt, Germany). Standard biochemical and hematological analyses were performed in a COBAS^® 8000 (Roche Diagnostics, Basel, Switzerland) and a Sysmex XN-1000^TM (Sysmex GmbH) automated analyzers.

PON1 activity was measured as the rate of hydrolysis of phenylacetate at 280 nm, in a 9 mM Tris-HCl buffer, pH 8.0, and supplemented with 0.9 mM CaCl2, as previously reported. PON1 can hydrolyze multiple substrates, but we chose phenylacetate because it is not toxic, the assay is simple, and is little influenced by PON1 gene polymorphisms [19 (link)]. PON1 concentration was analyzed with the Human PON1 ELISA kit from Elabscience^® (Houston, TX, USA). The concentrations of CCL2, interleukin (IL)-10, and IL-6, were analyzed with ABTS ELISA Development kits (Peprotech, London, UK). Galectin-3 was quantified with the Human Galectin-3 Quantikine ELISA Kit (R&D Systems^®, Minneapolis, MN, USA). Angiotensin II was measured with the Angiotensin II EIA kit from Sigma-Aldrich (St. Louis, MO, USA). ACE2 concentration was analyzed with the DuoSet^® ELISA Development System (R&D Systems^®, Minneapolis, MN, USA). Standard biochemical and hematological analyses were performed in a COBAS^® 8000 (Roche Diagnostics, Basel, Switzerland) and a Sysmex XN-1000^TM (Sysmex GmbH, Norderstedt, Germany), respectively.