Luminex bead based multiplex detection system

Blood samples were obtained from patients with TBI. The anticoagulated blood was centrifuged (1500 rpm, 15 min) to obtain the serum for ELISA analysis. Commercial ELISA Kits (ml037372, mlbio, RAG018R, BioVendor R&D, and NBP3-08117, Novus) were used to test and validate serum irisin levels, and the irisin-specific Luminex bead-based multiplex detection system (Merck Millipore) was further employed to detect human serum irisin. Detailed irisin assays were performed according to the kit instructions.

Peripheral venous blood samples were collected between 6 and 7 a.m. after overnight fasting, kept at room temperature for clotting and centrifuged at 3,000 × g for 15 min to obtain serum. Serum fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), creatinine (Cre) and blood urea nitrogen (BUN) levels were checked by an automatic chemistry analyzer (Beckman Coulter, Inc., Brea, CA, USA). The levels of the adipokines and myokines leptin, apelin, fractalkine, BDNF, LIF, myostatin, FABP3, irisin, FSTL1, oncostatin M, FGF21 and musclin in the serum were measured by an adipokine- and myokine-specific Luminex bead-based multiplex detection system (Merck Millipore, Darmstadt, Germany).