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Once the lysate had been loaded, the column was washed with 20 column volumes (CV) wash buffer, and the His-tagged TRFH domain was eluted using a linear gradient from 0 to 100% elution buffer (50 mM Tris-HCl pH 8.0, 500 mM NaCl, 1 mM TCEP, 300 mM imidazole) over 25 CV

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