Human agrin elisa kit

Cell culture supernatants were collected from each sample at 21 DIV (n = 6/group) and processed as mentioned earlier. Regular triculture media was used as control group. Human Agrin ELISA kit (Abcam, ab216945) was used to quantify Agrin levels following manufacturer’s protocol.

Blood samples were collected in Gel Clot Activator tubes (368969, BD Diagnostic, Oxford, UK) and centrifuged at 3000 rpm for 10 min, and then the serum was collected. Quantification of the circulating serum levels of the c-terminal agrin fragment (CAF), brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4) was performed by kits in duplicates. The adopted kits were the Human Agrin ELISA kit (ab216945, Abcam, Cambridge, UK), Human BDNF ELISA kit (ab212166, Abcam, Cambridge, UK) and Human NT-4 ELISA kit (EHNTF4, Invitrogen, Waltham, MA, USA), respectively. The dilutions for the sample were 1:4 for human agrin, 1:20 for human BDNF and no dilution for human NT-4. The NT-4 level was not detectable in 5 out of 11 subjects at any time point. Quantification was performed by a standard curve with the reconstituted standard provided by the kits and analysed using linear regression or with polynomials, depending on the best fit, as suggested by the guidelines. The absorbance, in accordance with the kit protocol, has been measured by a microplate ELISA reader (Tecan, Infinite F50, Kawasaki, Japan).