Human pentraxin 3 tsg 14 quantikine elisa kit

All biomarkers were analysed through commercially available kits. PTX3 was quantified in plasma samples through an enzyme-linked immunosorbent assay (ELISA) kit (Human Pentraxin 3/TSG-14 Quantikine ELISA Kit, R&D Systems, Minnesota, USA). Three classical inflammatory biomarkers were evaluated in serum samples: high-sensitivity (hs) CRP by immunoturbidimetry (Cardiac C-Reactive Protein (Latex) High Sensitive assay, Roche Diagnostics, Basel, Switzerland), IL-6, and TNF-α (Human IL-6 Quantikine HS and Human TNF-alpha Quantikine HS, R&D Systems). Lipid profile, including total cholesterol, triglycerides, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), was performed using laboratorial routine procedures (Cobas Integra 400 Plus autoanalyser; Roche Diagnostics, Basel, Switzerland); plasma oxidized LDL (oxLDL) and serum adiponectin, and leptin levels were determined through ELISA kits (Oxidized LDL ELISA Kit, Mercodia AB, Uppsala, Sweden; Human Total Adiponectin/Acrp30 Quantikine ELISA Kit and Human Leptin Quantikine ELISA Kit, R&D Systems). Levels of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) and serum tissue inhibitor of metalloproteinase-1 (TIMP-1) (Human proBNP and Human TIMP1 ELISA kits, Abcam, Cambridge, UK) were assessed as cardiac and renal fibrosis markers, respectively.

The amount of PTX3 protein released into supernatants was measured using the Human Pentraxin 3/TSG-14 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA).

A Human Pentraxin 3/TSG-14 Quantikine ELISA Kit (R&D Systems, Oxfordshire, UK) was used to measure PTX3 levels; the assay was carried out as per manufacturer’s protocol and read using an EnVision plate reader (Perkin-Elmer, MA, USA). The lower limit of detection for PTX3 is 0.31 ng/mL. Readings taken at 560 nm wavelengths were subtracted from the readings taken at 450 nm to correct for any optical imperfections within the plate. The average optical density from the negative control was subtracted from each well to remove any background noise present. All standards, negatives, and samples were run in duplicate on each plate. The standards were plotted against the optical density value. The R² value was >0.97.