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The pellet was resuspended in 200 µl of 8 M urea with 10 mM Tris (pH 8), incubated at 60° C for 30 min with shaking and centrifuged for 15 min at 18,000 g for protein quantification of the supernatant with the DC Protein Assay (Bio-Rad Laboratories)

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Every vial must be discarded after one use, even if water remains inside