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 |