ELISA: CXCL11/I-TAC Antibody (10C6) [NBP2-22533] - Sandwich ELISA of Human ITAC was performed using the Thermo Scientific ELISA Kit by coating a blank 96-well microtiter plate with 100ul per well of Human ITAC moncolonal antibody in duplicate at 1, 3, 4, 5, 7, and 9ug/ml in DPBS and incubating for 12-18 hours at 4C. The plate was aspirated and blocked with 300ul per well of 4% BSA and 5% sucrose in DPBS for 1 hour at room temperature. Human ITAC recombinant protein at 50ul per well was added in duplicate at 4000, 1600, 640, 256, 102.4, 40.96, and 0 pg/ml for 2 hours at room temperature along with 50ul of Human ITAC biotinylated monoclonal antibody in all applicable wells at 0.2ug/ml for 2 hours at room temperature. The plate was washed with ELISA Wash Buffer and incubated with 100ul per well of Streptavidin-HRP in all test wells at 1:80,000 dilution for 1 hour at room temperature and then washed and incubated with 100ul per well of TMB substrate for 30 minutes at room temperature in the dark. The plate was stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550nm.