View source for Team:UiOslo Norway/Experiments/Ni-NTA Affinity Chromatography

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<h1>Ni-NTA Affinity Chromatography</h1>
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  <li><p>Equilibriated the Ni-NTA with <i> E. coli </i> raw extract by rolling for 40 minutes.</p></li>
  <li><p>Transfer mixture to an Econo-Column® Chromatography Columns (BIORAD).</p></li>
  <li><p>Wash with 100 mL 10 mM imidazolebuffer and collect the fraction.</p></li>
  <li><p>Wash with 30 mL 50 mM imidazolebuffer and collect the fraction.</p></li>
  <li><p>Elute with 300 mM imidazolebuffer and collect the fraction.</p></li>
  <li><p>Check the obtained fractions by SDS-Page.</p></li>
  <p><b>Imidazole buffer</b></br>
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50 mM 		Tris-HCl pH 8 </br>
100 mM	NaCl</br>
10 mM  		beta-Mercaptoethanol</br>
10 mM         Imidazole </br>
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