Principle of Assay
Human Capicua ELISA Kit (A350444) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Capicua in serum, plasma, cell culture supernatant, cell or tissue lysate, 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 Capicua binds to the immobilized antigen. Following incubation, unbound components are removed by washing and a biotinylated antigen detection reagent is added to the wells. This reagent binds the captured Capicua to form an antigen-antibody-antigen sandwich complex. After further incubation and washing, HRP-Streptavidin conjugate is added to the wells. TMB substrate solution is then 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 Capicua captured in each well. The concentration of Capicua is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
