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Endocr Pract 2003, 9: 417-470

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The next day, acetone was removed post-centrifugation at 17,000 g at 4 C, washed twice with 80% ice-cold acetone, and resuspended in digestion buffer consisting of 3 M urea and 100 mM HEPES at pH 8.0 such that the final protein concentration was 1 g/L

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