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

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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

iGEM UiOslo 2015 is sponsored by:

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