Recipe

IMAC guanidine binding buffer

5 mM imidazole

1 mM 2-Mercaptoethanol

6 M Guanidine-HCl

20 mM Tris-HCl (pH 8.0)

Weigh out the guanidine, add imidazole and Tris-HCl (pH 8.0), 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 at room temperature overnight. Adjust the pH, add H2O to the final volume, and filter the solution. The pH adjustment must be done at room temperature. Add 2-mercaptoethanol immediately before use.

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  1. doi:10.1101/pdb.rec10051 Cold Spring Harb Protoc 2006: pdb.rec10051-

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