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. Author manuscript; available in PMC: 2022 May 21.
Published in final edited form as: J Vis Exp. 2021 May 21;(171):10.3791/62075. doi: 10.3791/62075

Table 1:

Composition of buffers, Ni-NTA denaturing and native purifications and Ion exchange low and high salt buffers.

Denaturing buffers compositions Native buffers compositions
Lysis buffer 10 mM NaH2PO4, 10 mM Tris, 6 M GuHCl, 10 mM imidazole, pH 8.0 20 mM Tris, 300 mM NaCl, 10 mM imidazole, pH 8.0
Wash buffer 10 mM NaH2PO4, 10 mM Tris, 6 M GuHCl, 20 mM imidazole, pH 8.0 20 mM Tris, 300 mM NaCl, 20 mM imidazole, pH 8.0
Elution buffer 10 mM NaH2PO4, 10 mM Tris, 6 M GuHCl, 0.2 M acetic acid, pH 3.0 20 mM Tris, 300 mM NaCl, 250 mM imidazole, pH 8.0
Buffer composition
High salt 5 mM Tris Acid, 5 mM Tris Base, pH 8.1, 4 M NaCl
Low salt 5 mM Tris Acid, 5 mM Tris Base, pH 8.1