Mouse high sensitivity ifn α elisa kit

An enzyme-linked immunosorbent assay (ELISA) was used to measure the concentrations of IFN-α and IFN-β in the spinal cord. Mouse high-sensitivity IFN-α ELISA kit and IFN-β ELISA kit were purchased from PBL Assay Science. Spinal tissues were homogenized in a lysis buffer containing protease and phosphatase inhibitors. Tissue samples were centrifuged at 12,500 ×g for 10 min and the supernatant was collected. BCA Protein Assay (Pierce) was employed to determine protein concentrations. For each reaction in a 96-well plate, 100 μg of proteins of samples were used. All ELISA experiments followed the manufacturer’s protocol. The optical densities of samples were measured using an ELISA plate reader (Bio-Rad) and the levels of IFN-α and IFN-β were calculated using the standard curves and ELISA results were reported normalized to protein concentration (pg/mg spinal tissue).

Mouse high-sensitivity IFN-α ELISA kit (42115-1) and IFN-β ELISA kit (42410-1) were purchased from PBL Assay Science. Mouse CTX-I ELISA kit (AC-06F1) and mouse PINP ELISA kit (AC-33F1) were purchased from Immunodiagnostic Systems. ELISA was performed using culture medium, serum, and bone marrow lysates. Serum was obtained from whole blood that was collected from a submandibular vein via facial vein puncture, coagulated for 30 min at room temperature, followed by centrifugation (2000 × g for 10 min, 4 °C) and collection of the supernatant (serum). Bone marrow was homogenized in a lysis buffer containing protease inhibitors at 4 °C for 30 min. ELISA was conducted in accordance with the manufacturer’s instructions. A standard curve was performed for each experiment.