IMAC urea binding buffer
To prepare 200 ml, add 2 ml 2 M imidazole, 20 ml 5 M NaCl, and 4 ml 1 M Tris to 96.1 g urea and H2O to 90% of the final volume. Heat the container, which is cooled by the solubilization process of the denaturant, to room temperature in a water bath at 45ºC while stirring. Avoid high temperatures. The denaturant will dissolve within minutes. Alternatively, stir overnight at room temperature. Adjust the pH, add H2O to adjust the final volume to 200 ml, and filter the solution. The pH adjustment must be done at room temperature. Add 14 µl of 2-mercaptoethanol immediately before use.
