Useful in Sandwich ELISA for Quantitative Detection of Antigen. Aliquot 0.1ml per well of the 1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml, 15.6pg/ml rat TNF alpha standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of rat cell culture supernates, serum or plasma(heparin , EDTA) to each empty well. It is recommended that each rat TNF alpha standard solution and each sample be measured in duplicate.