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

• HOME
• PROJECT
  • Description
  • Results
  • Achievements
• TEAM
  • Team
  • Attributions
  • Acknowledgement
• NOTEBOOK
  • Notebook
  • Experiments & Protocols
• PARTS
  • Team Parts
  • Basic Parts
• HUMAN PRACTICES
  • Human Practices
  • Collaborations
• SAFETY

Ni-NTA Affinity Chromatography

Back to Protocols

• 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