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3, set up calibration wells, sample diluent wells, blank wells and sample wells, and add different concentrations of calibration wells to each calibration well50L, add sample diluent to the sample diluent well50L, no addition is made to the blank wells, and the sample wells are added with the sample to be tested50LBiotin antibody was added to each well except the blank well.100uL, cover the reaction plate with a sealing film,37Incubate in a water bath or incubator away from light60min