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

 
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   <li><p>Elute with 300 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>
 
   <li><p>Check the obtained fractions by SDS-Page.</p></li>
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   <p><b>Imidazole buffer</b></br>
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50 mM Tris-HCl pH 8 </br>
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100 mM NaCl</br>
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10 mM  beta-Mercaptoethanol</br>
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10 mM        Imidazole </br>
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<!---------------sponsors------------------------->
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<center><h1>iGEM UiOslo 2015 is sponsored by:</h1>
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<hr style="height:2px;border:none;color:black;background-color:black;" />
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<img src="https://static.igem.org/mediawiki/2015/a/ac/UiOslo_Statoil.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/e/e0/UiOslo_uio_logo.png" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/0/08/UiOslo_Enova1.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/e/eb/UiOslo_GATC-Biotech.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/4/4a/UiOslo_IDT.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/d/d9/UiOslo_Norwegian_Biochemical_Society.jpg" Height="60px" hspace="15">
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<img src="https://static.igem.org/mediawiki/2015/f/f0/UiOslo_SnapGene.jpg" Height="60px" hspace="15">
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</center>
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<hr style="height:2px;border:none;color:black;background-color:black;" />
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<!--------end sponsors---------------------------->
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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: