Sauer:Purification of His-tagged proteins/Native prep

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Contents

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

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