https://eln.excellgen.com/index.php?action=history&feed=atom&title=2115_2203_with_chaperones 2115 2203 with chaperones - Revision history 2026-10-18T07:48:35Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2115_2203_with_chaperones&diff=11918&oldid=prev Paul.xu at 21:35, 28 May 2014 2014-05-28T21:35:22Z <p></p> <p><b>New page</b></p><div>== Use chaperones to improve Flp expression&lt;br&gt; ==<br /> <br /> 5/27/2014&lt;br&gt; <br /> <br /> Test chaperone on -yFLP expression&lt;br&gt; <br /> <br /> [[Image:May272014.jpg]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> {| width=&quot;832&quot; border=&quot;1&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | Lane&lt;br&gt; <br /> | Name&lt;br&gt; <br /> | Protein Name&lt;br&gt; <br /> | Clone ID&lt;br&gt; <br /> | Host&lt;br&gt; <br /> | Induction&lt;br&gt; <br /> | Purification&lt;br&gt; <br /> | sample&lt;br&gt;<br /> |-<br /> | 1&lt;br&gt; <br /> | rowspan=&quot;5&quot; | <br /> pT7A3-iFLP <br /> <br /> 9.76 / 52.8 &lt;br&gt; <br /> <br /> | rowspan=&quot;5&quot; | iFLP &lt;br&gt; &lt;br&gt; <br /> | rowspan=&quot;5&quot; | 2115&lt;br&gt; &lt;br&gt; <br /> | <br /> 15 ml,&lt;br&gt; <br /> <br /> Tuner(DE3)-pG-KJE8 <br /> <br /> | <br /> 0.2% arabinose+5ng/ml Tetracycline&lt;br&gt; <br /> <br /> 0.3 mM IPTG&amp;nbsp; 30oC o/n <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> S&lt;br&gt; <br /> <br /> |-<br /> | 2&lt;br&gt; <br /> | <br /> 1 5 ml, <br /> <br /> Tuner(DE3)-pGro7 &lt;br&gt; <br /> <br /> | <br /> 0.2% arabinose+&lt;br&gt; <br /> <br /> 0.3 mM IPTG 30oC o/n <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> S&lt;br&gt; <br /> <br /> |-<br /> | 3&lt;br&gt; <br /> | <br /> 15 ml, <br /> <br /> Tuner(DE3)-pKJE7 &lt;br&gt; <br /> <br /> | <br /> 0.2% arabinose+&lt;br&gt; <br /> <br /> 0.3 mM IPTG 30oC o/n <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> S <br /> <br /> |-<br /> | 4&lt;br&gt; <br /> | <br /> 15 ml, <br /> <br /> Tuner(DE3)-pGTf2&lt;br&gt; <br /> <br /> | <br /> 5ng/ml Tetracycline&lt;br&gt; <br /> <br /> 0.3 mM IPTG 30oC o/n <br /> <br /> | <br /> Ni&amp;nbsp; beads <br /> <br /> | <br /> S <br /> <br /> |-<br /> | 5&lt;br&gt; <br /> | 15 ml, <br /> Tuner(DE3)-pTf16&lt;br&gt; <br /> <br /> | <br /> 0.2% arabinose+&lt;br&gt; <br /> <br /> 0.3 mM IPTG 30oC o/n <br /> <br /> | Ni beads &lt;br&gt; <br /> | S&lt;br&gt;<br /> |-<br /> | 6&lt;br&gt; <br /> | Fermentas PageRuler Plus&lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> | <br /> &lt;br&gt; <br /> <br /> |-<br /> | 7&lt;br&gt; <br /> | rowspan=&quot;5&quot; | pT7A3-iFLP <br /> 9.76 / 52.8 &lt;br&gt; <br /> <br /> | rowspan=&quot;5&quot; | iFLP&lt;br&gt; <br /> | rowspan=&quot;5&quot; | 2115&lt;br&gt; <br /> | &amp;nbsp; 15 ml, <br /> Tuner(DE3)-pG-KJE8 <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> | 0.2% arabinose+5ng/ml Tetracycline <br /> 0.3 mM IPTG 30oC o/n&lt;br&gt; <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> p <br /> <br /> |-<br /> | 8&lt;br&gt; <br /> | &amp;nbsp;1 5 ml, <br /> Tuner(DE3)-pGro7 <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> | 0.2% arabinose+ <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | Ni beads&lt;br&gt; <br /> | <br /> p&lt;br&gt; <br /> <br /> |-<br /> | 9&lt;br&gt; <br /> | &amp;nbsp; 15 ml, <br /> Tuner(DE3)-pKJE7 <br /> <br /> &lt;br&gt; <br /> <br /> | 0.2% arabinose+ <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | <br /> Ni beads <br /> <br /> | p<br /> |-<br /> | 10&lt;br&gt; <br /> | 15 ml, <br /> Tuner(DE3)-pGTf2 <br /> <br /> &lt;br&gt; <br /> <br /> | 5ng/ml Tetracycline <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | Ni beads &lt;br&gt; <br /> | p&lt;br&gt;<br /> |-<br /> | 11&lt;br&gt; <br /> | 15 ml, <br /> Tuner(DE3)-pTf16&lt;br&gt; <br /> <br /> | 0.2% arabinose+ <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | Ni beads&lt;br&gt; <br /> | p&lt;br&gt;<br /> |-<br /> | 12&lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt;<br /> |}<br /> <br /> [[Image:May2720141.jpg]]&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> {| width=&quot;832&quot; border=&quot;1&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot;<br /> |-<br /> | Lane&lt;br&gt; <br /> | Name&lt;br&gt; <br /> | Protein Name&lt;br&gt; <br /> | Clone ID&lt;br&gt; <br /> | Host&lt;br&gt; <br /> | Induction&lt;br&gt; <br /> | Purification&lt;br&gt; <br /> | sample&lt;br&gt;<br /> |-<br /> | 1&lt;br&gt; <br /> | rowspan=&quot;3&quot; | <br /> pCold-MBP-Tat-NLS-iFLP-6H 9.03 / 96.9&lt;br&gt;&lt;br&gt; <br /> <br /> | rowspan=&quot;3&quot; | MBP-Tat-NLS-iFLP-6H&lt;br&gt; &lt;br&gt; <br /> | rowspan=&quot;3&quot; | 2203&lt;br&gt; <br /> | <br /> 15 ml,&lt;br&gt; <br /> <br /> Tuner(DE3)-pG-KJE8 <br /> <br /> | <br /> 0.2% arabinose+5ng/ml Tetracycline&lt;br&gt; <br /> <br /> 0.3 mM IPTG&amp;nbsp; 30oC o/n <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> S&lt;br&gt; <br /> <br /> |-<br /> | 2&lt;br&gt; <br /> | <br /> 1 5 ml, <br /> <br /> Tuner(DE3)-pGro7 &lt;br&gt; <br /> <br /> | <br /> 0.2% arabinose+&lt;br&gt; <br /> <br /> 0.3 mM IPTG 30oC o/n <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> S&lt;br&gt; <br /> <br /> |-<br /> | 3&lt;br&gt; <br /> | <br /> 15 ml, <br /> <br /> Tuner(DE3)-pKJE7 &lt;br&gt; <br /> <br /> | <br /> 0.2% arabinose+&lt;br&gt; <br /> <br /> 0.3 mM IPTG 30oC o/n <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> S <br /> <br /> |-<br /> | 4&lt;br&gt; <br /> | Fermentas PageRuler Plus&lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | <br /> &lt;br&gt; <br /> <br /> | <br /> &lt;br&gt; <br /> <br /> |-<br /> | 5&lt;br&gt; <br /> | rowspan=&quot;7&quot; | <br /> &lt;br&gt; pCold-MBP-Tat-NLS-iFLP-6H <br /> <br /> 9.03 / 96.9&lt;br&gt;&lt;br&gt; <br /> <br /> | rowspan=&quot;7&quot; | &lt;br&gt; MBP-Tat-NLS-iFLP-6H&lt;br&gt; <br /> | rowspan=&quot;7&quot; | &lt;br&gt; 2203 &lt;br&gt; <br /> | 15 ml, <br /> Tuner(DE3)-pGTf2 <br /> <br /> &lt;br&gt; <br /> <br /> | 5ng/ml Tetracycline <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | Ni beads &lt;br&gt; <br /> | S&lt;br&gt;<br /> |-<br /> | 6&lt;br&gt; <br /> | 15 ml, <br /> Tuner(DE3)-pTf16&lt;br&gt; <br /> <br /> | 0.2% arabinose+ <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | <br /> Ni beads&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> | <br /> s&lt;br&gt; <br /> <br /> |-<br /> | 7&lt;br&gt; <br /> | &amp;nbsp; 15 ml, <br /> Tuner(DE3)-pG-KJE8 <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> | 0.2% arabinose+5ng/ml Tetracycline <br /> 0.3 mM IPTG 30oC o/n&lt;br&gt; <br /> <br /> | <br /> Ni beads <br /> <br /> | <br /> p <br /> <br /> |-<br /> | 8&lt;br&gt; <br /> | &amp;nbsp;1 5 ml, <br /> Tuner(DE3)-pGro7 <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> | 0.2% arabinose+ <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | Ni beads&lt;br&gt; <br /> | <br /> p&lt;br&gt; <br /> <br /> |-<br /> | 9&lt;br&gt; <br /> | &amp;nbsp; 15 ml, <br /> Tuner(DE3)-pKJE7 <br /> <br /> &lt;br&gt; <br /> <br /> | 0.2% arabinose+ <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | <br /> Ni beads <br /> <br /> | p<br /> |-<br /> | 10&lt;br&gt; <br /> | 15 ml, <br /> Tuner(DE3)-pGTf2 <br /> <br /> &lt;br&gt; <br /> <br /> | 5ng/ml Tetracycline <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | Ni beads &lt;br&gt; <br /> | p&lt;br&gt;<br /> |-<br /> | 11&lt;br&gt; <br /> | 15 ml, <br /> Tuner(DE3)-pTf16&lt;br&gt; <br /> <br /> | 0.2% arabinose+ <br /> 0.3 mM IPTG 30oC o/n &lt;br&gt; <br /> <br /> | Ni beads&lt;br&gt; <br /> | p&lt;br&gt;<br /> |-<br /> | 12&lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt; <br /> | &lt;br&gt;<br /> |}<br /> <br /> &lt;br&gt; Conclusion: Chaperones do not enhance protein expression. Protein is also insoluble&lt;br&gt;</div> Paul.xu