Sauer:Purification of His-tagged proteins/Denaturing prep
From OpenWetWare
This is a general protocol for purification of His6-tagged proteins under denaturing conditions. Note that you may need to modify this protocol to fit your specific needs.
Buffers
Lysis buffer
100 mM NaH2PO4
10 mM Tris base
6 M guanidine hydrochloride
10 mM imidazole
Adjust to pH 8
Wash buffer 1
100 mM NaH2PO4
150 mM NaCl
8 M urea
20 mM imidazole
Adjust to pH 8
Wash buffer 2
50 mM NaH2PO4
500 mM NaCl
20 mM imidazole
Adjust to pH 8
Elution buffer
50 mM NaH2PO4
500 mM NaCl
250 mM imidazole
Adjust to pH 8
