Principle of Assay
Human Peroxiredoxin 1/PAG ELISA Kit (A77180) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Peroxiredoxin 1/PAG in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for Peroxiredoxin 1/PAG. Standards and samples are added to the wells, where Peroxiredoxin 1/PAG binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody specific for Peroxiredoxin 1/PAG is added to the wells. The detection antibody binds the captured Peroxiredoxin 1/PAG 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 Peroxiredoxin 1/PAG captured in each well. The concentration of Peroxiredoxin 1/PAG is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
