http://openwetware.org/index.php?title=Pulse-chase_protein_production&feed=atom&action=history 2013-05-25T07:04:02Z Revision history for this page on the wiki MediaWiki 1.13.2 http://openwetware.org/index.php?title=Pulse-chase_protein_production&diff=153909&oldid=prev 2007-09-25T11:32:07Z

<p></p> <table style="background-color: white; color:black;"> <col class='diff-marker' /> <col class='diff-content' /> <col class='diff-marker' /> <col class='diff-content' /> <tr valign='top'> <td colspan='2' style="background-color: white; color:black;">←Older revision</td> <td colspan='2' style="background-color: white; color:black;">Revision as of 11:32, 25 September 2007</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 1:</td> <td colspan="2" class="diff-lineno">Line 1:</td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">I don't know what this heading is supposed to mean</del>.&nbsp; <del class="diffchange diffchange-inline">Labeling? Production?</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">To determine the amount of protein generated during a given period of time, whether bulk protein in the cell or a particular protein from a gel or other separation technique, grow the cell with radiolabelled amino acids for that time, then resuspend the cells in medium lacking the radioactive amino acids</ins>.&nbsp; <ins class="diffchange diffchange-inline">At time points afterwards, beginning with immediately after resuspension, isolate protein from the cells and measure its concentration (say by a Bradford assay) and the radioactivity of your preparation.&nbsp; The radioactivity per unit of protein immediately after the end of labelling tells you how strongly the protein was labelled during that period.&nbsp; The decrease in radioactivity per unit of protein over time tells you how fast the protein is turned over.</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">When hearing this title, the first thing that would come to my mind is to use a synthetic gene regulatory network, perhaps a puslation or oscillator device, to control expression of a heterologous protein. Maybe such a system can </del>be <del class="diffchange diffchange-inline">developed </del>to <del class="diffchange diffchange-inline">help overcome inclusion body formation; a system which checks </del>the <del class="diffchange diffchange-inline">level </del>of <del class="diffchange diffchange-inline">chaperons before producing more recombinant </del>protein. <del class="diffchange diffchange-inline">Thats my wild guess of what Pulse-chase could mean -Devin</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">It may </ins>be <ins class="diffchange diffchange-inline">important </ins>to <ins class="diffchange diffchange-inline">take </ins>the <ins class="diffchange diffchange-inline">supernatant </ins>of <ins class="diffchange diffchange-inline">the culture and measure the radioactivity of its protein fraction to know about secreted </ins>protein.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">---</del></div></td><td colspan="2">&nbsp;</td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2">&nbsp;</td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">I am fairly certain that pulse-chase protein production refers to experiments known as pulse-labeling. This is when you subject your organism to an environmental stimulus (hot, cold, IPTG, etc.), you wait some amount of time, then introduce a radio-labeled amino acid (typically methionine). So the pulse is the stimulus and the chase is the label such as hot methionine. - Lon</del></div></td><td colspan="2">&nbsp;</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:Needs attention]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:Needs attention]]</div></td></tr> <!-- diff generator: internal 2013-05-25 07:04:02 --> </table>

Madhadron http://openwetware.org/index.php?title=Pulse-chase_protein_production&diff=117075&oldid=prev 2007-05-16T23:42:29Z

<p></p> <table style="background-color: white; color:black;"> <col class='diff-marker' /> <col class='diff-content' /> <col class='diff-marker' /> <col class='diff-content' /> <tr valign='top'> <td colspan='2' style="background-color: white; color:black;">←Older revision</td> <td colspan='2' style="background-color: white; color:black;">Revision as of 23:42, 16 May 2007</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 2:</td> <td colspan="2" class="diff-lineno">Line 2:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>When hearing this title, the first thing that would come to my mind is to use a synthetic gene regulatory network, perhaps a puslation or oscillator device, to control expression of a heterologous protein. Maybe such a system can be developed to help overcome inclusion body formation; a system which checks the level of chaperons before producing more recombinant protein. Thats my wild guess of what Pulse-chase could mean -Devin</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>When hearing this title, the first thing that would come to my mind is to use a synthetic gene regulatory network, perhaps a puslation or oscillator device, to control expression of a heterologous protein. Maybe such a system can be developed to help overcome inclusion body formation; a system which checks the level of chaperons before producing more recombinant protein. Thats my wild guess of what Pulse-chase could mean -Devin</div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">---</ins></div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">I am fairly certain that pulse-chase protein production refers to experiments known as pulse-labeling. This is when you subject your organism to an environmental stimulus (hot, cold, IPTG, etc.), you wait some amount of time, then introduce a radio-labeled amino acid (typically methionine). So the pulse is the stimulus and the chase is the label such as hot methionine. - Lon</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:Needs attention]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Category:Protocol]] [[Category:Needs attention]]</div></td></tr> <!-- diff generator: internal 2013-05-25 07:04:02 --> </table>

LonChubiz http://openwetware.org/index.php?title=Pulse-chase_protein_production&diff=115708&oldid=prev 2007-05-09T18:40:47Z

<p></p> <table style="background-color: white; color:black;"> <col class='diff-marker' /> <col class='diff-content' /> <col class='diff-marker' /> <col class='diff-content' /> <tr valign='top'> <td colspan='2' style="background-color: white; color:black;">←Older revision</td> <td colspan='2' style="background-color: white; color:black;">Revision as of 18:40, 9 May 2007</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 2:</td> <td colspan="2" class="diff-lineno">Line 2:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>When hearing this title, the first thing that would come to my mind is to use a synthetic gene regulatory network, perhaps a puslation or oscillator device, to control expression of a heterologous protein. Maybe such a system can be developed to help overcome inclusion body formation; a system which checks the level of chaperons before producing more recombinant protein. Thats my wild guess of what Pulse-chase could mean -Devin</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>When hearing this title, the first thing that would come to my mind is to use a synthetic gene regulatory network, perhaps a puslation or oscillator device, to control expression of a heterologous protein. Maybe such a system can be developed to help overcome inclusion body formation; a system which checks the level of chaperons before producing more recombinant protein. Thats my wild guess of what Pulse-chase could mean -Devin</div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">[[Category:Protocol]] [[Category:Needs attention]]</ins></div></td></tr> <!-- diff generator: internal 2013-05-25 07:04:02 --> </table>

Reshma P. Shetty http://openwetware.org/index.php?title=Pulse-chase_protein_production&diff=80476&oldid=prev 2006-10-21T02:35:18Z

<p></p> <table style="background-color: white; color:black;"> <col class='diff-marker' /> <col class='diff-content' /> <col class='diff-marker' /> <col class='diff-content' /> <tr valign='top'> <td colspan='2' style="background-color: white; color:black;">←Older revision</td> <td colspan='2' style="background-color: white; color:black;">Revision as of 02:35, 21 October 2006</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 1:</td> <td colspan="2" class="diff-lineno">Line 1:</td></tr> <tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>I don't know what this heading is supposed to mean.&nbsp; Labeling? Production?</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>I don't know what this heading is supposed to mean.&nbsp; Labeling? Production?</div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"></ins></div></td></tr> <tr><td colspan="2">&nbsp;</td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;">When hearing this title, the first thing that would come to my mind is to use a synthetic gene regulatory network, perhaps a puslation or oscillator device, to control expression of a heterologous protein. Maybe such a system can be developed to help overcome inclusion body formation; a system which checks the level of chaperons before producing more recombinant protein. Thats my wild guess of what Pulse-chase could mean -Devin</ins></div></td></tr> <!-- diff generator: internal 2013-05-25 07:04:02 --> </table>

DeviNcAmenares http://openwetware.org/index.php?title=Pulse-chase_protein_production&diff=11696&oldid=prev 2005-10-06T15:19:52Z

<p></p> <table style="background-color: white; color:black;"> <col class='diff-marker' /> <col class='diff-content' /> <col class='diff-marker' /> <col class='diff-content' /> <tr valign='top'> <td colspan='2' style="background-color: white; color:black;">←Older revision</td> <td colspan='2' style="background-color: white; color:black;">Revision as of 15:19, 6 October 2005</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 1:</td> <td colspan="2" class="diff-lineno">Line 1:</td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>&#160;</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">I don't know what this heading is supposed to mean.&nbsp; Labeling? Production?</ins></div></td></tr> <!-- diff generator: internal 2013-05-25 07:04:02 --> </table>

Smoore http://openwetware.org/index.php?title=Pulse-chase_protein_production&diff=7300&oldid=prev 2005-06-15T23:04:12Z

<p></p> <table style="background-color: white; color:black;"> <col class='diff-marker' /> <col class='diff-content' /> <col class='diff-marker' /> <col class='diff-content' /> <tr valign='top'> <td colspan='2' style="background-color: white; color:black;">←Older revision</td> <td colspan='2' style="background-color: white; color:black;">Revision as of 23:04, 15 June 2005</td> </tr> <tr><td colspan="2" class="diff-lineno">Line 1:</td> <td colspan="2" class="diff-lineno">Line 1:</td></tr> <tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Please fill me in...</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>&#160;</div></td></tr> <!-- diff generator: internal 2013-05-25 07:04:02 --> </table>

Skosuri http://openwetware.org/index.php?title=Pulse-chase_protein_production&diff=1410&oldid=prev 2005-06-07T19:45:04Z

<p></p> <p><b>New page</b></p><div>Please fill me in...</div>

Igem2005