Principle of Assay
Guinea Pig Ovalbumin Specific IgE ELISA Kit (A302600) is a 4 hour indirect Enzyme-Linked Immunosorbent Assay (iELISA) designed for the in vitro quantitative determination of guinea pig Ovalbumin Specific IgE in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with assay-specific antigen. Standards and samples are added to the wells, where Ovalbumin Specific IgE present in the samples binds to the immobilized antigen. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody is added to the wells. This detection antibody binds the captured Ovalbumin Specific IgE to form an indirect immune complex. After further incubation and washing, TMB substrate solution is added to visualize the HRP enzymatic 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.
