The 12 PPP reference specimens were received from BD. An aliquot from each of 12 serum samples was divided into a portion to be labeled with NHS-digoxigenin and a portion to be labeled with NHS-biotin (Molecular Probes). The digoxigenin-labeled samples were pooled, and equal amounts of the pool were transferred to each of the biotin-labeled samples. Each labeled protein solution was supplemented with nonfat milk to a final concentration of 3%, Tween-20 to a final concentration of 0.1%, and 1 x PBS to yield a final serum dilution of 1:100.
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