Description
This assay employs an antibody specific for Anti-human TNF Alpha coated on a 96-well plate. Standards and samples are pipetted into the wells and TNF Alpha present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human TNF Alpha antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of TNF Alpha bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm