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> | ||
| − | + | <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> | ||
| + | </ul> | ||
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<!---------------sponsors-------------------------> | <!---------------sponsors-------------------------> | ||
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| − | <center><h1>iGEM UiOslo 2015 | + | <center><h1>iGEM UiOslo 2015 is sponsored by:</h1> |
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| + | <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/2015/0/08/UiOslo_Enova1.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"> | <img src="https://static.igem.org/mediawiki/2015/f/f0/UiOslo_SnapGene.jpg" Height="60px" hspace="15"> | ||
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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:
