Editing MS 3 20 2017 (section)

== <span class="mw-headline" id="Regeneration_of_Ni_column"><span class="mw-headline" id="Regeneration_of_Ni_column"><span class="mw-headline" id="Regeneration_of_Ni_column">Regeneration of Ni column</span></span></span> ==

1. Wash Ni&nbsp;column with 5 CV of water.

2. Wash column with 3 CV of 100 mM EDTA, 500 mM NaCl.

3. Wash the column with 5 CV of H2O.

4. Recharge the column with 1 column volume of 100 mM NiSO4, or NiCl2.

5. Wash the column with 10 column volumes of H2O.

6. Equilibrate with 3 volumes of a suitable buffer (''e.g.,&nbsp;''Buffer A).







'''Buffer A'''

20 mM Tris-HCl, pH 7.9~8.0

500 mM NaCl

0.1 mM EDTA

0.1% Tween 20

0.1% Nonident-P40

<br/>'''Ni Washing Buffer'''

20 mM Tris-HCl, pH 7.9~8.0

800 mM NaCl

10 mM imidazole&nbsp;





'''Tris-HCl Buffer&nbsp;'''

20 mM Tris-HCl, pH 7.9~8.0





'''Buffer B&nbsp;(Low Salt)'''

20 mM Tris-HCl, pH 7.9~8.0

100 mM NaCl

150&nbsp;mM imidazole&nbsp;

5% glycerol





'''Dilution Buffer'''

20 mM Tris-HCl, pH 7.9~8.0, optional: 5% glycerol





'''Q HP Washing Buffer'''

20 mM Tris-HCl, pH 7.9~8.0

50 mM KCl

5% Trehalose





'''Q HP Elution&nbsp;Buffer'''

20 mM Tris-HCl, pH 7.9~8.0

1 M KCl

5% Trehalose





'''Buffer E'''

20 mM HEPES (pH 8.3) or Tris-HCl, pH 8.0&nbsp;





'''Buffer F'''

20 mM HEPES (pH 8.0) or Tris-HCl, pH 7.9&nbsp;

50 mM KCl.&nbsp;





'''Buffer G'''

20 mM HEPES or Tris-HCl, pH 7.9 or pH 8.0

500 mM KCl.





'''Storage Buffer'''

20 mM HEPES, pH 7.5

100 mM KCl

0.1 mM EDTA

1 mM dithiothreitol

50% glycerol.
[[Category:My ELN]] [[Category:Protein Expression]] [[Category:QC]]