Pulse-chase protein production - Revision history

Madhadron at 11:32, 25 September 2007

Madhadron — Tue, 25 Sep 2007 11:32:07 GMT

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-	~~I don't know what this heading is supposed to mean~~. ~~Labeling? Production?~~	+	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. 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. The radioactivity per unit of protein immediately after the end of labelling tells you how strongly the protein was labelled during that period. The decrease in radioactivity per unit of protein over time tells you how fast the protein is turned over.

-	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~~	+	It may be important to take the supernatant of the culture and measure the radioactivity of its protein fraction to know about secreted protein.

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-	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

	[[Category:Protocol]] [[Category:Needs attention]]		[[Category:Protocol]] [[Category:Needs attention]]

LonChubiz at 23:42, 16 May 2007

LonChubiz — Wed, 16 May 2007 23:42:29 GMT

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	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		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
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		+	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

	[[Category:Protocol]] [[Category:Needs attention]]		[[Category:Protocol]] [[Category:Needs attention]]

Reshma P. Shetty at 18:40, 9 May 2007

Reshma P. Shetty — Wed, 09 May 2007 18:40:47 GMT

←Older revision		Revision as of 18:40, 9 May 2007
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	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		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
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		+	[[Category:Protocol]] [[Category:Needs attention]]

DeviNcAmenares at 02:35, 21 October 2006

DeviNcAmenares — Sat, 21 Oct 2006 02:35:18 GMT

←Older revision		Revision as of 02:35, 21 October 2006
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	I don't know what this heading is supposed to mean. Labeling? Production?		I don't know what this heading is supposed to mean. Labeling? Production?
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		+	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

Smoore at 15:19, 6 October 2005

Smoore — Thu, 06 Oct 2005 15:19:52 GMT

←Older revision	Revision as of 15:19, 6 October 2005
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-	+	I don't know what this heading is supposed to mean. Labeling? Production?

Skosuri at 23:04, 15 June 2005

Skosuri — Wed, 15 Jun 2005 23:04:12 GMT

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-	~~Please fill me in...~~	+

Igem2005 at 19:45, 7 June 2005

Igem2005 — Tue, 07 Jun 2005 19:45:04 GMT

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