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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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This measure was the consequence to the devastating consequences for the ecosystem by the widespread use of DDT, which continuously grew due to it's strong insecticidal properties

Many of them are used off-label, meaning outside the original approved purpose