Nickel NTA purification of 6-his Proteins under Denaturing Conditions
Dominik Haudenschild February 20, 2001 at UCD
Concentrate Protein
Concentrate protein from Supernatant of cells using centricon, or alternatively TCA precipitate
Bind protein to Ni-NTA Agarose (Qiagen)
- Add 250µl protein to 9.5ml Binding Buffer
8M Urea, 10mM Tris, 0.1M Sodium Phosphate pH 8.0
- Add 250µl Ni-NTA Agarose slurry
- Allow to bind for 20 minutes on rocker platform