Principle of Assay
Human Frataxin ELISA Kit (A303264) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Frataxin in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for Frataxin. Standards and samples are added to the wells, where Frataxin binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody specific for Frataxin is added to the wells. The detection antibody binds the captured Frataxin to form an antibody-antigen-antibody sandwich 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 Frataxin captured in each well. The concentration of Frataxin is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
