Team:UiOslo Norway/Experiments/Ni-NTA Affinity Chromatography

Ni-NTA Affinity Chromatography

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  • Equilibriated the Ni-NTA with E. coli raw extract by rolling for 40 minutes.

  • Transfer mixture to an Econo-Column® Chromatography Columns (BIORAD).

  • Wash with 100 mL 10 mM imidazolebuffer and collect the fraction.

  • Wash with 30 mL 50 mM imidazolebuffer and collect the fraction.

  • Elute with 300 mM imidazolebuffer and collect the fraction.

  • Check the obtained fractions by SDS-Page.

  • Imidazole buffer

    50 mM Tris-HCl pH 8
    100 mM NaCl
    10 mM beta-Mercaptoethanol
    10 mM Imidazole