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After washing with HCl (step 3), the two cartridges were connected in series and the rest of the procedure was performed.
Extraction was carried on in bead mill with two tungsten beads and the rest of the procedure was the same as for plasma samples.
For ELISA, plates were coated as for panning and the rest of the procedure was performed as previously described [ 19].
The stored urine was allowed to thaw at room temperature and then diluted 1 : 100, and the rest of the procedure was according to the manufacturer's instructions.
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We also created control samples by omitting only the primary antibodies and treating the rest of the procedures similarly.
This procedure was repeated 100 times to include unused samples, and for each iteration the reduced data set was run through the rest of the procedure.
The rest of the procedure was performed as described for S100A4 and osteopontin.
The rest of the procedure remains the same as OMP.
The rest of the procedure required no operator input for the actual segmentation procedure.
The rest of the procedure was identical to that in Experiment 1.
The rest of the procedure was as per the miniprep manual (Qiagen).
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