Principle of Assay
Human NEK2 ELISA Kit (A78269) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human NEK2 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 NEK2 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 NEK2 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 NEK2 captured in each well. The concentration of NEK2 is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
