Human neurotrophin 3 elisa kit

Levels of NT3 protein in the serum were assessed using a Human Neurotrophin-3 ELISA Kit (#ab100615 Abcam), which can detect vector-derived and endogenous NT3. Whole blood was centrifuged (7200 g, 25̊C), the serum removed and stored at −80 °C. ELISA was performed as per the manufacturer's instructions and the plate read immediately at 450 nm using a BMG LabTech FLUOStar (Omega). NT3 levels were normalised to the total amount of protein extracted from each sample determined by a Bicinchoninic acid assay (BCA) assay (#71285–3, Novagen, Millipore). Serum samples were diluted 1:150 in PBS, and a four-parameter standard curve was generated (above r > 0.99).

NT-3 concentration of plasma samples of patients and healthy controls was quantified by sandwich enzyme-linked immunosorbent assay (ELISA), using a commercial kit (Human Neurotrophin 3 ELISA Kit, ab100615, Lote No. GR3229491-1, Abcam, Cambridge, UK), according to the manufacturer’s instructions. Tests were performed in duplicate, and an automated microplate reader (Biochrom ASYS UVM 340, Cambridge, UK) measured the optical density at 450 nm with Mikrowin 2000 software (Berthold Technologies, Germany). Plasma total proteins were measured by a bicinchoninic acid assay (BCA, Pierce Chemicals, Rockford, IL, USA).