https://eln.excellgen.com/index.php?action=history&feed=atom&title=2381TRX-iFLP_PKAalpha_purification_2014_12_05 2026-10-18T02:19:42Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2381TRX-iFLP_PKAalpha_purification_2014_12_05&diff=13712&oldid=prev 2014-12-04T22:10:31Z

<p>Created page with &quot;2014/12/1--12/5 &lt;br&gt; <a href="/index.php?title=File:2181_PKAapurification.jpg" title="File:2181 PKAapurification.jpg">Image:2181 PKAapurification.jpg</a>&lt;br&gt; 2381 pT7A3-TRX-Tat-NLS-iFlp-6H 8L Nico21(DE3)plysN when OD600=2.0 use 0.2 mM IPTG RT induce overnight, 40g&lt;br&gt; a...&quot;</p> <p><b>New page</b></p><div>2014/12/1--12/5 <br /> <br /> &lt;br&gt; [[Image:2181 PKAapurification.jpg]]&lt;br&gt; <br /> <br /> 2381 pT7A3-TRX-Tat-NLS-iFlp-6H 8L Nico21(DE3)plysN when OD600=2.0 use 0.2 mM IPTG RT induce overnight, 40g&lt;br&gt;<br /> <br /> add lysis buffer, freeze and thaw, then ultracentrifuge, &lt;br&gt;<br /> <br /> supernatant: load to yellow Ni column----elute ----diluted load to sp column--elute&lt;br&gt;<br /> <br /> pellet : use urea and&amp;nbsp; guanidine hydrochloride to dissolve, load to yellow Ni ----elute&lt;br&gt;<br /> <br /> lane1-4: 2381 protein eluted from sp column&lt;br&gt;<br /> <br /> lane5: 2381flowthrough from sp column&lt;br&gt;<br /> <br /> lane6-8: 2381 protein eluted from Ni column&lt;br&gt;<br /> <br /> PKAa&lt;br&gt;<br /> <br /> lane10: PKAa lysate&lt;br&gt;<br /> <br /> lane11: PKAa lysate SUPERNATANT purified with 75 ul Ni beads&lt;br&gt;<br /> <br /> lane12:PKAa lysate PELLET purified with 75 ul Ni beads&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> [[Category:Protein_Purification]]</div>

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