Sauer:Purification of His-tagged proteins/Native prep: Difference between revisions
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=Introduction= | =Introduction= | ||
This is a general protocol for purification of His<sub>6</sub>-tagged proteins under native conditions. Note that you may need to modify this protocol to fit your specific needs. | This is a general protocol for purification of His<sub>6</sub>-tagged proteins under native conditions. Note that you may need to modify this protocol to fit your specific needs. This protocol is a modified version of that provided with Ni<sup>2+</sup>-NTA resin from [http://www1.qiagen.com/SelectCountry.aspx Qiagen]. | ||
=Buffers= | =Buffers= | ||
===Lysis buffer=== | ===Lysis buffer=== | ||
Latest revision as of 16:08, 9 February 2006
Introduction
This is a general protocol for purification of His6-tagged proteins under native conditions. Note that you may need to modify this protocol to fit your specific needs. This protocol is a modified version of that provided with Ni2+-NTA resin from Qiagen.
Buffers
Lysis buffer
50 mM NaH2PO4
500 mM NaCl
10 mM imidazole
Adjust to pH 8
Wash buffer
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
