Principle of Assay
Rat Ovalbumin Specific IgE ELISA Kit (A303777) is a capture Enzyme-Linked Immunosorbent Assay (cELISA) designed for the in vitro quantitative determination of rat Ovalbumin Specific IgE in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with a capture antibody specific for Ovalbumin Specific IgE. Standards and samples are added to the wells, where Ovalbumin Specific IgE present in each sample binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an assay-specific detection reagent is added to the wells. This detection reagent binds the captured Ovalbumin Specific IgE to form a capture immune complex. After further incubation and washing, TMB substrate solution is added to visualize the enzyme-mediated reaction. This produces a blue-coloured product that changes to yellow following addition of acidic stop solution. The intensity of the yellow colour is directly proportional to the amount of Ovalbumin Specific IgE captured in each well. The concentration of Ovalbumin Specific IgE is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
