Difference between revisions of "Team:UiOslo Norway/Experiments/Ni-NTA Affinity Chromatography"

 
(One intermediate revision by the same user not shown)
Line 21: Line 21:
 
10 mM        Imidazole </br>
 
10 mM        Imidazole </br>
 
</ul>
 
</ul>
 +
<div class="clear"></div>
 +
<!---------------sponsors------------------------->
 +
 +
 +
 +
<center><h1>iGEM UiOslo 2015 is sponsored by:</h1>
 +
<hr style="height:2px;border:none;color:black;background-color:black;" />
 +
<img src="https://static.igem.org/mediawiki/2015/a/ac/UiOslo_Statoil.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/e/e0/UiOslo_uio_logo.png" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/0/08/UiOslo_Enova1.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/e/eb/UiOslo_GATC-Biotech.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/4/4a/UiOslo_IDT.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/d/d9/UiOslo_Norwegian_Biochemical_Society.jpg" Height="60px" hspace="15">
 +
<img src="https://static.igem.org/mediawiki/2015/f/f0/UiOslo_SnapGene.jpg" Height="60px" hspace="15">
 +
</center>
 +
<hr style="height:2px;border:none;color:black;background-color:black;" />
 +
 +
<!--------end sponsors---------------------------->
 +
 +
  
 
</body>
 
</body>
 
</html>
 
</html>

Latest revision as of 12:20, 18 September 2015

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

iGEM UiOslo 2015 is sponsored by: